AU2023202271A1 - RNAi approach for crop pest protection - Google Patents

RNAi approach for crop pest protection Download PDF

Info

Publication number
AU2023202271A1
AU2023202271A1 AU2023202271A AU2023202271A AU2023202271A1 AU 2023202271 A1 AU2023202271 A1 AU 2023202271A1 AU 2023202271 A AU2023202271 A AU 2023202271A AU 2023202271 A AU2023202271 A AU 2023202271A AU 2023202271 A1 AU2023202271 A1 AU 2023202271A1
Authority
AU
Australia
Prior art keywords
dsrna
sexta
insect
seq
plant
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
AU2023202271A
Inventor
Bala P. VENKATA
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Donald Danforth Plant Science Center
Original Assignee
Donald Danforth Plant Science Center
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Donald Danforth Plant Science Center filed Critical Donald Danforth Plant Science Center
Priority to AU2023202271A priority Critical patent/AU2023202271A1/en
Publication of AU2023202271A1 publication Critical patent/AU2023202271A1/en
Pending legal-status Critical Current

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7088—Compounds having three or more nucleosides or nucleotides
    • A61K31/713—Double-stranded nucleic acids or oligonucleotides
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
    • A01N63/60—Isolated nucleic acids
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/113—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
    • C12N15/8216—Methods for controlling, regulating or enhancing expression of transgenes in plant cells
    • C12N15/8218—Antisense, co-suppression, viral induced gene silencing [VIGS], post-transcriptional induced gene silencing [PTGS]
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
    • C12N15/8241—Phenotypically and genetically modified plants via recombinant DNA technology
    • C12N15/8261—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield
    • C12N15/8271—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance
    • C12N15/8279—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance for biotic stress resistance, pathogen resistance, disease resistance
    • C12N15/8286—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance for biotic stress resistance, pathogen resistance, disease resistance for insect resistance
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2310/00—Structure or type of the nucleic acid
    • C12N2310/10—Type of nucleic acid
    • C12N2310/14—Type of nucleic acid interfering N.A.
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
    • Y02A40/10—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in agriculture
    • Y02A40/146—Genetically Modified [GMO] plants, e.g. transgenic plants

Abstract

Provided herein is the identification of insect RNAi target genes (MIGGS-IRTG) involved in gut microbial clearance and containment induced by microbes ingested during feeding. In certain aspects, the target gene is critical for insect immune responses and abundantly expressed in the midgut. Also provided are examples of a novel biotechnology for devising pesticidal RNAi approaches.

Description

RNAi APPROACH FOR CROP PEST PROTECTION
BACKGROUND
[0001] Insect pests are detrimental to crop production and human health throughout the world and insect control can in some instances consume between 10-25% of a country's gross national product (GNP). (http World Wide Web internet site "fao.org/3/a-avOl3e.pdf'). In the U.S., annual loss due to crop pests is estimated to exceed $120 billion USD/year. (Polaszek A. (1998) Wallingford, UK: CABI. 530 pp.).
[0002] Within crop pests, Lepidoptera are the most detrimental insect pests of cereal crop cultivation. Chemical control is often expensive, inefficient, and can be associated with negative environmental consequences. Host plant resistance is an attractive option but impeded by lack of robust Lepidoptera resistant germplasm (http World Wide Web internet site "cnbc.com/2015/05/08/insects-feast-on-plants-endangering-crops-and-costing-billions.html").
[0003] Since 1996, commercialization of crop plants genetically engineered to produce Bacillus thuringiensis(Bt) insecticidal proteins have resulted in efficient pest control, increased yield, reduced insecticidal use, and enhanced farmer profits. (Khan ZR, et al. (2014). Philos. Trans. R Soc. Lond Biol Sci. 369: 1639).
[0004] Consequently, the cumulative area planted with Bt crops worldwide reached greater than 1 billion acres during 2011. Within the U.S., Bt Corn, Bt Soybean, and Bt Cotton .0 accounted for 90% of all the total corn, soybean and cotton acres during 2013 (Tabashnik BE, et al. (2013). Nat. Biotech. 31: 510-521). However, evolution of field resistance against Bt in lepidopteran pests raises potential concerns about the sustainability of this approach. (Campagne P., et al. (2013) PLoS ONE 8(7): e69675. Doi:10.1371). That is further exacerbated by loss of resistance against pyramided Bt traits as well (https World Wide Web internet site dt npf.com/agriculture/web/ag/news/article/2016/08/10/rootworm-resistance-pyramided-bt.html).
SUMMARY
[0005] Provided herein is an isolated double stranded RNA (dsRNA) molecule comprising a nucleic acid sequence complementary to about 200 to 1000 contiguous nucleotides of a target gene sequence-wherein the target gene is a MIGGS-IRTG as defined herein-involved in gut microbe clearance and/or containment induced by microbes ingested during feeding and/or active feeding. In certain aspects, the target gene is critical for insect immune responses and certain aspects provide that it is abundantly expressed in the midgut. In certain aspects, the target gene sequence includes at least one of the protein coding region, the 5' UTR region, the 3' UTR region, and any combination thereof, of a target gene. Further, certain aspects provide that the dsRNA molecule silences the target gene when ingested by an insect. In certain aspects, the target gene is a type 1 MIGGS RNAi target or a type 2 MIGGS RNAi target as defined elsewhere herein. In certain aspects, the target gene is a pattern recognition receptor (PRR) class gene or an insect midgut structural component gene. In certain aspects, the target gene is expressed abundantly in a midgut specific manner during active feeding.
[0006] In certain aspects, a dsRNA molecule disclosed anywhere herein comprises two annealed complementary RNA strands. In certain aspects, said dsRNA molecule comprises a single RNA strand comprising an inversely repeated sequence with a spacer in between, wherein the single RNA strand can anneal to itself to form a hairpin loop structure.
[0007] In certain aspects, a dsRNA molecule disclosed anywhere herein comprises a nucleic acid sequence complementary to about 200 to 1000 contiguous nucleotides of the protein coding region of the target gene sequence. In certain aspects, said dsRNA molecule comprises a nucleic acid sequence complementary to about 200 to 1000 contiguous nucleotides of the 5' UTR region or the 3' UTR region of the target gene sequence. In certain aspects, said dsRNA molecule comprises a nucleic acid sequence complementary to a contiguous region comprising at least about 50%, 60%, 70%, 80%, 90%, 95%, 9 8 %, 9 9 %, or 100% of the length of the target gene sequence protein coding region, 5' UTR region, or 3' UTR region. In certain aspects, said dsRNA molecule comprises a nucleic acid sequence complementary to about 200 to 650 contiguous nucleotides of a target gene sequence.
[0008] Certain aspects of this disclosure are drawn to a target gene selected from the group consisting of M sexta-Hemolin (MsHEM), M. sexta-Serine proteinase homolog 3 (MsSPH-3), M. sexta-Peptidoglycan recognition protein 2 (MsPGRP2), M sexta-Beta-1, 3 glycan-recognition protein 2 (MspGRP2), M. sexta-Relish family protein 2A (MsREL2A), M sexta-Dorsal (MsDor), M. sexta-Sptzle (MsSPZ1A), M. sexta-Toll receptor (MsTOLL), M sexta-Scolexin A (MsSCA1), M. sexta-Hemolymph proteinase 18 (MsHP18), M sexta Transferrin (MsTRN), M. sexta-Arylphorin beta subunit (MsARP), M sexta-Chymotrypsinogen like protein 1 (MsCTL1), M sexta-Valine Rich Midgut Protein (MsVMP1), M. sexta-Imd (MsImd), M sexta-FADD (MsFADD), M. sexta-Dredd (MsDRD), M. sexta-Relish F (MsReIF),
M. sexta-Cdc42 (MsCdc42), M sexta-Dsorl (MsDsorl), M sexta-Fos (MsFos), M sexta-Jra (MsJra), M. sexta-Caudal (MsCAD), M sexta-Atg8 (MsAtg8), M sexta-Atgl3 (MsAtgl3), M sexta-IAP1(MsIAP1), M sexta-Chitin synthase 2 (MsChs2), M sexta-Beta-1 tubulin (MsTub), M. sexta-Beta fructofuranosidase 1 (MsSucl), and orthologs thereof In certain aspects, the target gene is selected from the group consisting ofM. sexta-Hemolin (MsHEM), M sexta-Serine proteinase homolog 3 (MsSPH-3), M sexta-Peptidoglycan recognition protein 2 (MsPGRP2), M sexta-Beta-1, 3-glycan-recognition protein 2 (MsGRP2), M. sexta-Relish family protein 2A (MsREL2A), M sexta-Dorsal (MsDor), M. sexta-Spatzle (MsSPZ1A), M. sexta-Toll receptor (MsTOLL), M. sexta-Scolexin A (MsSCA1), M. sexta-Hemolymph proteinase 18 (MsHP18), M sexta-Transferrin (MsTRN), M sexta-Arylphorin beta subunit (MsARP), M sexta Chymotrypsinogen-like protein 1 (MsCTL1), M sexta-Valine Rich Midgut Protein (MsVMIP1), M. sexta-Imd (MsImd), M sexta-FADD (MsFADD), M. sexta-Dredd (MsDRD), M sexta-Relish F (MsReIF), M sexta-Cdc42 (MsCdc42), M sexta-Dsorl (MsDsorl), M sexta-Fos (MsFos), M. sexta-Jra (MsJra), M. sexta-Caudal (MsCAD), M sexta-Atg8 (MsAtg8), M sexta-Atgl3 (MsAtgl3), M sexta-IAP1(MsIAP1), M. sexta-Chitin synthase 2 (MsChs2), M. sexta-Beta-1 tubulin (MspTub) andM. sexta-Beta fructofuranosidase 1 (MsSuc1).
[0009] Also provided herein is an isolated double stranded RNA (dsRNA) molecule comprising a nucleic acid sequence complementary to about 200 to 1000 contiguous nucleotides of a target gene sequence, wherein the target gene comprises a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 1-14, 16-29, 31-70, 71-75, 76-88, 89-105, and 106 110. In certain aspects, the target gene sequence includes at least one of the protein coding region, the 5' UTR region, the 3' UTR region, and any combination thereof, of a target gene. Further, certain aspects provide that the dsRNA molecule silences the target gene when ingested by an insect.
[0010] In certain aspects disclosed herein, the target gene is sequence selected from the group consisting of: i) SEQ ID NOs: 1-9, 11, 14, 31, 39, 43, 44, and 71-75.
[0011] In certain aspects disclosed herein, the target gene is sequence selected from the group consisting of: ii) SEQ ID NOs: 3, 4, and 43. In certain aspects, the dsRNA molecule causes impeded growth, developmental progression, and/or mortality and the like of TH, DMB, and FAW in an orthologous manner.
[0012] In certain aspects disclosed herein, the target gene is sequence selected from the group consisting of: iii) SEQ ID NOs: 76, 77, 80, 81, 85, 87, and 88. In certain aspects, the dsRNA molecule causes impeded growth, developmental progression, and/or mortality and the like of DBM. Further, in certain aspects, the DBM is a Bt resistant strain.
[0013] In certain aspects disclosed herein, the target gene is sequence selected from the group consisting of: iv) SEQ ID NOs: 89, 92, 96, 101, 103, and 105. In certain aspects, the dsRNA molecule causes impeded growth, developmental progression, and/or mortality and the like of FAW.
[0014] In certain aspects disclosed herein, the target gene is sequence selected from the group consisting of: v) SEQ ID NOs: 107-110. In certain aspects, the dsRNA molecule caused impeded growth, developmental progression, and/or mortality and the like of RFB.
[0015] In certain aspects of the dsRNA molecule disclosed above, the dsRNA molecule comprises two annealed complementary RNA stands. In certain aspects, the dsRNA molecule comprises a single RNA strand comprising an inversely repeated sequence with a spacer in between and where the single RNA strand can anneal to itself to form a hairpin loop structure.
[0016] In certain of the dsRNA molecule disclosed above, the dsRNA molecule comprises a nucleic acid sequence complementary to about 200 to 1000 contiguous nucleotides of the protein coding region of the target gene sequence. In certain aspects, the dsRNA molecule comprises a nucleic acid sequence complementary to a contiguous region comprising at least about 50%, 60%, 70%, 80%, 90%, 95%, 98%, 99%, or 100% of the length of a sequence selected from the group consisting of SEQ ID NOs: 1-14, 16-29, 31-70, 71-75, 76-88, 89-105, and 106 110. In certain aspects, the dsRNA molecule comprises a nucleic acid sequence complementary to a contiguous region comprising at least about 50%, 60%, 70%, 80%, 90%, 95%, 98%, 99%, or 100% of the length of the target gene sequence protein coding region, 5' UTR region, or 3' UTR region.
[0017] In certain aspects disclosed herein the dsRNA molecule comprises a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 111-119, 120-126, 127-135, and 136-139. In certain aspects, the dsRNA is a fragment of at least about 200 nucleotides thereof In certain aspects, i) the isolated dsRNA molecule comprising a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 110-119, or the fragment thereof, causes impeded growth, developmental progression, and/or mortality and the like of TH; ii) the isolated dsRNA molecule comprising a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 120-126, or the fragment thereof, causes impeded growth, developmental progression, and/or mortality and the like of DBM; iii) the isolated dsRNA molecule comprising a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 127-135, or the fragment thereof, causes impeded growth, developmental progression, and/or mortality and the like of FAW; or iv) the isolated dsRNA molecule comprising a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 136-139, or the fragment thereof, causes impeded growth, developmental progression, and/or mortality and the like of RFB.
[0018] In certain of any of the above aspects, the dsRNA molecule can form siRNA. Certain aspects provide for an isolated siRNA molecule derived from the processing of said dsRNA molecule.
[0019] Certain further aspects provide of an insecticidal composition comprising an isolated dsRNA molecule or an siRNA molecule disclosed anywhere herein, and a synthetic carrier or microbial conduit. In certain aspects, a microorganism has a natural capacity or is engineered to produce and/ or deliver dsRNA to increase its bioavailability and/or biostability for causing RNA interference including but not restricted to plant growth promoting organisms, normal commensal and/or symbiotic microorganisms associated with the target insect pest or parasites and/or natural enemies of the target pest or pest target host or host cultivation range etc. from an insect or parasite and/or natural enemies of the target pest engineered or identified from natural populations containing microbial conduit to produce and/or deliver dsRNA and/or drive the transmission of such microbial conduits into natural populations of insect pests as a control option. In certain aspects of an insecticidal composition disclosed herein, the dsRNA molecule is .0 conjugated with the synthetic carrier.
[0020] Certain aspects are also drawn to a recombinant DNA construct encoding a dsRNA molecule disclose anywhere herein. In certain aspects, the recombinant DNA construct comprising a gene silencing sequence comprising about 200 to 1000 contiguous nucleotides of a target gene sequence. In certain aspects, the target gene is a MIGGS-IRTG, as defined herein, involved in gut microbe clearance and/or containment induced by microbes ingested during feeding and/or active feeding. In certain aspects, the target gene is critical for insect immune responses. In certain aspects, the target gene is abundantly expressed in the midgut. In certain aspects, said target gene sequence includes at least one of the protein coding region, the 5' UTR region, the 3' UTR region, and any combination thereof, of a target gene. In certain aspects, the target gene is a type I MIGGS RNAi target or a type 2 MIGGS RNAi target as described elsewhere herein. In certain aspects, the target gene is a pattern recognition receptor (PRR) class gene or an insect midgut structural component gene. In certain aspects, the target gene is expressed abundantly in a midgut specific manner during active feeding.
[0021] In any of the above aspects of a recombinant DNA construct, the gene silencing sequence comprises about 200 to 1000 contiguous nucleotides of the protein coding region of the target gene sequence. In certain aspects, the gene silencing sequence comprises about 200 to 1000 contiguous nucleotides of the 5' UTR region or the 3' UTR region of the target gene sequence. In certain aspects, the gene silencing sequence comprises at least 50%, 60%, 70%, 80%, 90%, 95%, 98%, 99%, or 100% contiguously of the length of target gene sequence protein coding region, 5' UTR region, or 3' UTR region. In certain aspects, the gene silencing sequence comprises about 200 to 650 contiguous nucleotides of the target gene sequence.
[0022] In any of the above aspects of a recombinant DNA construct, and as noted throughout this disclosure, in certain aspects, a target gene can be selected from the group consisting of M. sexta-Hemolin (MsHEM), M. sexta-Serine proteinase homolog 3 (MsSPH-3), M. sexta-Peptidoglycan recognition protein 2 (MsPGRP2), M sexta-Beta-1, 3-glycan-recognition protein 2 (MspGRP2), M sexta-Relish family protein 2A (MsREL2A), M. sexta-Dorsal (MsDor), M. sexta-Spatzle (MsSPZ1A), M sexta-Toll receptor (MsTOLL), M sexta-Scolexin A (MsSCA1), M. sexta-Hemolymph proteinase 18 (MsHP18), M sexta-Transferrin (MsTRN), M sexta-Arylphorin beta subunit (MsARP), M sexta-Chymotrypsinogen-like protein 1 (MsCTL1), M. sexta-Valine Rich Midgut Protein (MsVMP1), M sexta-Imd (MsImd), M. sexta-FADD .0 (MsFADD), M sexta-Dredd (MsDRD), M sexta-Relish F (MsReIF), M sexta-Cdc42 (MsCdc42), M. sexta-Dsorl (MsDsorl), M. sexta-Fos (MsFos), M sexta-Jra (MsJra), M. sexta Caudal (MsCAD1), M. sexta-Atg8 (MsAtg8), M. sexta-Atgl3 (MsAtgl3), M sexta IAP1(MsIAP1), M sexta-Chitin synthase 2 (MsChs2), M. sexta-Beta-1 tubulin (MspTub), M sexta-Beta fructofuranosidase 1 (MsSuc1), and orthologs thereof In certain aspects a target gene can be selected from the group consisting of M. sexta-Hemolin (MsHEM), M. sexta-Serine proteinase homolog 3 (MsSPH-3), M sexta-Peptidoglycan recognition protein 2 (MsPGRP2), M sexta-Beta-1, 3-glycan-recognition protein 2 (MspGRP2), M. sexta-Relish family protein 2A (MsREL2A), M sexta-Dorsal (MsDor), M. sexta-Spatzle (MsSPZ1A), M. sexta-Toll receptor (MsTOLL), M. sexta-Scolexin A (MsSCA1), M. sexta-Hemolymph proteinase 18 (MsHP18), M sexta-Transferrin (MsTRN), M sexta-Arylphorin beta subunit (MsARP), M sexta Chymotrypsinogen-like protein 1 (MsCTL1), M sexta-Valine Rich Midgut Protein (MsVMIP1), M. sexta-Imd (MsImd), M sexta-FADD (MsFADD), M. sexta-Dredd (MsDRD), M sexta-Relish
F (MsReIF), M sexta-Cdc42 (MsCdc42), M sexta-Dsorl (MsDsorl), M sexta-Fos (MsFos), M. sexta-Jra (MsJra), M. sexta-Caudal (MsCAD), M sexta-Atg8 (MsAtg8), M sexta-Atgl3 (MsAtgl3), M sexta-IAP1(MsIAP1), M. sexta-Chitin synthase 2 (MsChs2), M. sexta-Beta-1 tubulin (MspTub) andM. sexta-Beta fructofuranosidase 1 (MsSuc1).
[0023] Further aspects provide for a recombinant DNA construct comprising a gene silencing sequence comprising about 200 to 1000 contiguous nucleotides of a target gene sequence, wherein the target gene comprises a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 1-14, 16-29, 31-70, 71-75, 76-88, 89-105, and 106-110. In certain aspects, the target gene sequence includes at least one of the protein coding region, the 5' UTR region, the 3' UTR region, and any combination thereof, of a target gene.
[0024] In any of the above aspects of a recombinant DNA construct, the target gene sequence is selected from the group consisting of: i) SEQ ID NOs: 1-9, 11, 14, 31, 39, 43, 44, and 71-75; ii) SEQ ID NOs: 3, 4, and 43; iii) SEQ ID NOs: 76, 77, 80, 81, 85, 87, and 88; iv) SEQ ID NOs: 89, 92, 96, 101, 103, and 105; and v) SEQ ID NOs: 107-109, and 110. In certain aspects, the gene silencing sequence comprises about 200 to 1000 contiguous nucleotides of a sequence selected from the group consisting of SEQ ID NOs: 1-14, 16-29, 31-70, 71-75, 76-88, 89-105, and 106-110. In certain aspects, the gene silencing sequence comprises at least 50%, 60%, 70%, 80%, 90%, 95%, 98%, 99%, or 100% contiguously of a sequence selected from the group consisting of SEQ ID Nos: 1-14, 16-29, 31-70, 71-75, 76-88, 89-105, and 106-110. In .0 certain aspects, the gene silencing sequence comprises about 200 to 650 contiguous nucleotides of a sequence selected from the group consisting of SEQ ID NOs: 1-14, 16-29, 31-70, 71-75, 76 88, 89-105, and 106-110. In certain aspects, the gene silencing sequence is operably linked to one or more promoters for the expression of a dsRNA molecule that silences the target gene when ingested by an insect. In certain aspects, the construct is an expression vector. And, in certain aspects, the expression vector can target single or multiple insect RNAi target genes or chimeric RNAi target genes.
[0025] Certain aspects of the disclosure also provide for a host cell comprising the dsRNA molecule, the siRNA molecule, a polynucleotide encoding a dsRNA molecule, and/or the construct or a dsRNA encoding segment thereof disclose anywhere herein. In certain aspects, the host cell is a bacterial or plant cell or organelle. In certain aspects, the organelle is a plastid. In certain aspects, the host cell is a transgenic and/or transplastomic plant cell. In certain aspects, the hose cell expresses a dsRNA and/or produces an siRNA disclosed anywhere herein.
[0026] Certain aspects also provide for a transgenic and/or transplastomic plant comprising a dsRNA molecule, an siRNA molecule, a polynucleotide encoding the dsRNA, and/or a construct or a dsRNA encoding segment disclosed anywhere herein. In certain aspects, at least one cell of the plant expresses the dsRNA molecule and/or produces the siRNA molecule. Further, certain aspects provide for a seed, part, tissue, cell, or organelle of the above transgenic and/or transplastomic plant. In certain aspects, the seed, part, tissue, cell, or organelle comprises the dsRNA molecule and/or the siRNA molecule. In certain aspects, the organelle is a plastid.
[0027] Certain aspects provide for a method of silencing: (i) an insect immune response gene and/or (ii) an insect gene encoding for structural components of the insect midgut. In certain aspects, the method comprises providing for ingesting an isolated dsRNA molecule, an siRNA molecule, an insecticidal composition, a host cell, a transgenic and/or transplastomic plant, transplastomic plant and/or a seed, part, tissue, cell, or organelle as disclosed anywhere herein, to an insect.
[0028] Certain aspects provide for a method of protecting a plant from an insect pest of the plant. In certain aspects, the method comprises topically applying to a plant an isolated dsRNA molecule, an siRNA molecule, and/or an insecticidal composition disclosed anywhere herein, and providing the plant in the diet of the insect pest. In certain aspects, the dsRNA is topically applied by expressing the dsRNA in a microbe and topically applying the microbe onto the plant.
[0029] Certain aspects provide for a method of producing a plant resistance to a pest insect of said plant. In certain aspects, the method comprises transforming a plant with a polynucleotide encoding the dsRNA molecule and/or a construct or a dsRNA encoding segment thereof as disclosed anywhere herein, wherein the plant expresses a dsRNA molecule and/or produces an siRNA disclose anywhere herein.
[0030] Certain aspects provide for a method of improving crop yield. In certain aspects, the method comprises growing a population of crop plants transformed with a polynucleotide encoding a dsRNA molecule and/or a construct or a dsRNA encoding segment thereof wherein the plant expresses a dsRNA molecule and/or produces an siRNA molecule as discloses anywhere herein. In certain aspects, the population of transformed plants produces higher yields in the presence of pest insect infestation than a control population of untransformed plants.
[0031] Certain aspects provide for a method for producing a plant resistant against a pest insect of said plant. In certain aspects, the method comprises: a) transforming a plant cell and/or organelle with a polynucleotide encoding a dsRNA molecule and/or a construct or a dsRNA encoding segment thereof as disclosed anywhere herein; b) regenerating a plant from the transformed plant cell and/or organelle; and c) growing the transformed plant under conditions suitable for the expression of said double stranded RNA molecule, wherein said transformed plant of (c) is resistant to the plant pest insect compared to an untransformed plant.
[0032] In certain of any of the aforementioned aspects, the method the dsRNA is ingested by an actively feeding stage of the insect. In certain aspects, the ingestion of the dsRNA induces a melanotic response in the insect larvae. In certain aspects, the ingestion of the dsRNA results in perturbation of gut microbial homeostasis. In certain aspects, the ingestion of the dsRNA results in defective clearance of opportunistic microbes. In certain aspects, the ingestion of the dsRNA results in defective containment of gut microbes.
[0033] In certain of any of the aforementioned aspects, the silencing of the target gene results in reduced appetite and/or developmental defects resulting in incomplete development and/or mortality and/or decrease the reproductive success of the insect. In certain aspects, the reduced appetite and/or developmental defects and/or mortality and/or reduced reproductive fitness of the insect is observed after sustained feeding for at least 72 hours.
[0034] In certain of any of the aforementioned aspects, the insect is of the order Lepidoptera, Coleoptera, Hemiptera, Blattodea, or Diptera. In certain aspects, the insect is Manduca sexta (M sexta) (tobacco hornworm), Spodoptera frugiperda (fall armyworm), .0 Ostrinia nubilalis (European corn borer), Plutella xylostella (Diamondback moth), Leptinotarsa decemlineata Say (Colorado potato beetle), Diabrotica spp. (Corn rootworm complex), Tribolium castaneum (Red flour beetle), Popil/ajaponica(Japanese beetle), Agrilusplanipennis (Emerald ash borer), Diaphorina citri (Asian citrus psyllid), Cimex lectularius (Bed bug), a cockroach or termite, or insect pests such as mosquitoes and flies.
[0035] In certain of any of the aforementioned aspects, the plant host is selected from the group consisting of Zea mays L (corn), Sorghum bicolor (sorghum), Setaria italica (fox tail millet), Pennisetum glaucum (Pearl millet), Solanum tuberosum (potato), Oryza sativa (rice), Lycopersicon esculentum (tomato), Solanum melongena (eggplant), cultivars of the Brassica oleraceafamily, Citrussinensis (Orange), trees of the Oleaceaefamily, and crops of Rosaceae.
BRIEF DESCRIPTION OF THE DRAWINGS
[0036] Figure 1. Figure 1 shows schematic representation of bacterially ingestible dsRNA assay. (Kamath RS, et al. (2000) Genome Biol. 2: 1-10; Newmark et al. (2003) Proc. Natl. Acad. Sci. USA 100: 11861-11865).
[0037] Figure 2A-C. Figure 2A-C shows representative phenotypes of TH larvae. TH larvae exposed to bacterially (HT115 (DE3)) expressed dsRNA against MIGGS RNAi targets MsPGRP2 (A); MsVMPl(B); and negative control dsRNA against Cassava plant specific gene MeCAT I (C).
[0038] Figure 3A,B. Figure 3A,B shows feeding activity of TH larvae exposed to dsRNA against negative control MeCAT1 (A) and MIGGS RNAi target MsPGRP2 (B) containing bacterial (HT115 (DE3)) plates.
[0039] Figure 4. Figure 4 shows survival rates of healthy conventionally-reared (CR) and germ-free (GF) first instar TH larvae exposed to bacterially (HT115 (DE3)) expressed dsRNA against MIGGS RNAi targets MsPGRP2; MsVMP1 and negative control dsRNA against Cassava plant specific gene MeCATI. The differences observed using 4 replicates/treatment were statistically significant across all time points at a P value between P<0.001 to P<0.05.
[0040] Figure 5. Figure 5 shows percentage melanotic reaction of healthy conventionally-reared (CR) and germ-free (GF) first instar TH larvae exposed to bacterially (HT115 (DE3)) expressed dsRNA against MIGGS RNAi targets MsPGRP2; MsVMP1 and negative control dsRNA against Cassava plant specific gene MeCATI. The differences observed using 4 replicates/treatment were statistically significant across all time points at a P value between P<0.001 to P<0.05.
[0041] Figure 6. Figure 6 shows schematic representation of dsRNA producing L4440 vector containing coding sequence of Cassava CATI (MeCATI) and TH MIGGS RNAi targets MsPGRP2 and MsVMPl.
[0042] Figure 7. Figure 7 shows midgut-preferred expression of two TH MIGGS RNAi target genes MsHEM and MsSPH3 in comparison to the control gene RPS3. The control cDNA libraries were derived from conventionally reared larvae and treatment cDNA libraries were derived from TH larvae injected with 75 CFU of E. coi. The control and treatment larvae were used to isolate hemolymph fraction (HL), dissect midgut (MDG) to obtain rest of the body (RB). The HL, MDG and RB were used for RNA isolation and cDNA synthesis.
[0043] Figure 8. Figure 8 shows schematic representation of oral induction procedure for MIGGS RNAi target genes. The insect larvae were reared on induction media containing live gram-negative bacteria E. coli and lyophilized cell wall signatures from gram-positive bacteria and fungi, following a previously published protocol (Wang et al. (2006). J. Biol. Chem. 281(14): 9271-9278).
[0044] Figure 9A,B. Figure 9A,B shows expression of MIGGS RNAi target genes in TH larvae in response to feeding on induction media. Genes with immunity function (A,B) are induced (I) between 24-48 hours post larval exposure to induction media and mostly not detected in the absence of induction (U).
[0045] Figure 9C,D. Figure 9C,D also shows expression of MIGGS RNAi target genes in TH larvae in response to feeding on induction media. Genes with immunity function (C) are induced (I) between 24-48 hours post larval exposure to induction media and mostly not detected in the absence of induction (U). While, the genes essential for midgut structural integrity (D) are expressed under both conditions.
[0046] Figure 1OA-F. Figure 1OA-F shows representative phenotypes of TH larvae (initial size shown in A) exposed to bacterially expressed dsRNA against insecticidal MIGGS RNAi target genes MsToll2, MsSucl, and MspTub in comparison to negative (B) and positive (C) control treatment. The insecticidal activity is manifested as stunted growth and development, loss of appetite and melanotic reaction (D-F) in comparison to regular growth and development .0 observed with negative control treatment (B).
[0047] Figure 11. Figure 11 shows percentage mortality of TH larvae feeding on bacterially expressed dsRNA against insecticidal MIGGS RNAi targets MsToll2, MsSucl, and MspTub. The insecticidal MIGGS candidates confer statistically significant mortality that is comparable to positive control treatment (MsVATPaseE). Data is average of 3 replicates/treatment SEM at p< 0.001(***).
[0048] Figure 12A-C. Figure 12A-C shows the induction of MIGGS-IRTGS in DBM larvae feeding on induction media. The genes with immunity function (A-B) are induced (I) between 24-48 hours post larval exposure to induction media and mostly not detected in the absence of induction (U). While, the genes essential for midgut structural integrity (C) are expressed under both conditions.
[0049] Figure 13A-H. Figure 13A-H shows representative phenotypes of Bt resistant DBM larvae (initial size shown in A) exposed to bacterially expressed dsRNA against insecticidal DBM MIGGS RNAi targets PxPGRP2 (D), PxIMD (E), PxpGRP2 (F), PxCAC (G), and PxCHS1 (H) in comparison to positive (C) and negative control (B) treatment. The insecticidal activity is manifested as stunted growth and development, loss of appetite and melanotic reaction (D-H) in comparison to regular growth and development observed with negative control treatment (B).
[0050] Figure 14. Figure 14 shows percentage mortality of DBM larvae feeding on bacterially expressed dsRNA against insecticidal MIGGS RNAi targets PxPGRP2, PxIMD, PxPGRP2, PxCAC, and PxCHS1. The insecticidal MIGGS candidates confer statistically significant mortality that is comparable to positive control treatment (MsVATPaseE). Data is average of 3-replicates/treatment SEM at p< 0.001(***) p< 0.01(**).
[0051] Figure 15A,B. Figure 15A,B illustrates a sprayable RNAi set up using dsRNA against TH MIGGS targets MsPGRP2, Ms pGRP2, MsCHS2, and MsVMPl. One cm2 leaf discs from field soil grown tobacco plants (A) were drop inoculated with various concentrations of purified dsRNA against TH MIGGS RNAi targets (B). The bioassays were carried out for 5 days with 3 first instar larvae per well and leaf discs changed at the end of every 24 hours.
[0052] Figure 16. Figure 16 shows percentage mortality of TH larvae feeding on various concentrations of dsRNA against the core set of TH MIGGS RNAi targets MsPGPRP2, MspGRP2, MsCHS2, and MsVMP1. The leaf disc coated dsRNA causes significant mortality at 8 and 16 pg of dsRNA concentration. The leaf disc coated dsRNA against three core MIGGS .0 targets confers statistically significant mortality that is comparable to positive control treatment (MsVATPaseE). Data is average of 3-replicates/treatment SEM at p< 0.001(***); p< 0.01(**) and p 0.05(***).
[0053] Figure 17A-H. Figure 17A-H shows representative phenotypes of TH larvae (initial size shown in A and E) exposed to 8 and 16 jig of dsRNA against the core set of of MIGGS RNAi targets MsPGPRP2 (D), MspGRP2 (F), MsCHS2 (G), and MsVMP1 (H). The insecticidal activity is comparable to the positive control treatment (C) and manifested as stunted growth and development, loss of appetite and melanotic reaction (D,F-G) in contrast to the regular growth and development observed with negative control treatment (B).
[0054] Figure 18A-D. Figure 18A-D shows representative phenotypes of TH larvae feeding on leaves from transplastomic plants expressing dsRNA against core MIGGS targets MsPGRP2 (PTS-28-10 and PTS-28-7-B), MspGRP2 (PTS-26-19 and PTS-26-4-C), and dsCHS2 (PTS-27-3-1 and PTS-27-13-1-D). The TH larvae feeding on leaves expressing dsRNA against
MIGGS targets MsPGRP2 (B), MspGRP (C), and MsCHS2 (D) display stunted growth, development, loss of appetite and melanotic reaction in comparison to negative control PTS-27 13-2 (A).
[0055] Figure 18E. Figure 18E shows that the insecticidal activity of transplastomic lines is manifested as significant reduction in mean weights in comparison to negative control.
[0056] Figure 18F. Figure 18F shows that the transplastomic events confer significant mortality in comparison to negative control. The mortality rate was scored on a 0-3 score were 0, 1, 2 and 3 indicated < 0, 25, 50 or > 50 mortality respectively. Data is average of 6 replicates/treatment (N=24) SEM at p< 0.001(***); p< 0.01(**) and p< 0.05(*).
[0057] Figure 19. Figure 19 shows percentage mortality of Bt resistant DBM larvae feeding on various concentrations of dsRNA against the DBM orthologs of TH core MIGGS RNAi targets PxPGPRP2, PxpGRP2, PxpTUB, and PxCHS1. The leaf disc coated dsRNA causes significant mortality at 0.5-1 pg of dsRNA concentration. The leaf disc coated dsRNA against all core MIGGS targets confers statistically significant mortality between 0.5 g and 1 pg dsRNA dosage that is comparable to positive control treatment (MsVATPaseE). Data is average of 3 replicates/treatment SEM at p< 0.01(**) and p< 0.05(*).
[0058] Figure 20A-H. Figure 20A-H shows representative phenotypes of Bt resistant DBM larvae (initial size shown in A and E) exposed to 0.5-1 pg of dsRNA against the DBM orthologs of TH core MIGGS RNAi targets PxPGPRP2(D), PxpGRP2(F), PxpTUB(G), and .0 PxCHS1(H). The insecticidal activity is comparable to the positive control treatment (C) and manifested as stunted growth and development, loss of appetite and melanotic reaction (D,F-H) in contrast to the regular growth and development observed with negative control treatment (B).
[0059] Figure 21A-D. Figure 21A-D illustrates MIGGS-IRTG induction procedure in FAW feeding on wheat plants grown microbe rich field soil (A) and microbe depleted sterile surface (B). Ten first instar FAW larvae (C) were infested into each pot containing five wheat seedlings and contained using porous netting material (D). The larval samples were collected at various time points after infestation and used for pooled RNA-Seq approach to identify differential expressed transcripts in response to induction by the microbes in the filed soil.
[0060] Figure 22A,B. Figure 22A,B shows bi-clustering comparison of differential gene expression in FAW larvae feeding on microbe-depleted plants (A) in comparison to larvae feeding on microbe rich plants (B). RNA-Seq data indicated that plants growing on field soil caused preferential up-regulation of MIGGS pathway genes in FAW. In total, 100 differential expressed genes were identified, 30 of which were associated with MIGGS pathways. Notably, FAW orthologs of TH insecticidal targets including PGRP2, pGRP2 and IMD were captured in the data set, indicating clearly that MIGGS pathway genes are up regulated in response to insect feeding on plants exposed to soil microbiome.
[0061] Figure 23A-F. Figure 23A-F shows representative phenotypes of FAW larvae exposed to pure dsRNA against FAW orthologs of TH core MIGGS RNAi targets SFCHS2 (B), SFGRP2 (C), SFPGRP2 (D), and two novel MIGGS RNAi targets from RNA-Seq study SFRC (un-annotated-E) and C-type lectin-6 (SFCTL-F). Leaf discs coated with dsRNA causes reduced growth, development and loss of appetite in comparison to negative control treatment (A) when supplied at 8 and 16 pg of dsRNA concentration per leaf disc.
[0062] Figure 23G,H. Figure 23G,H illustrates from Figure 23A-F significant weight reduction (G) and mortality (H). The rates of mortality was scored on a 0-3 scale where 0, 1, 2 and 3 indicated < 0, 25, 50 or > 50 mortality respectively. The dsRNA treatments imposed caused statistically significant reduction in mean weights (G) that translated into significant rates of mortality (H) in comparison to negative control. Data is average of 3 replicates/treatment SEM at p< 0.001(***); p< 0.01(**) and p< 0.05(*).
[0063] Figure 24A,B. Figure 24 shows mean weights (A) of FAW larvae exposed to 16pg of pure dsRNA against newly discovered MIGGS RNAi targets from the RNA-Seq data. The rates of mortality was scored on a 0-3 score were 0, 1, 2 and 3 indicated <0, 25, 50 or > 50 .0 mortality respectively. The dsRNA treatments imposed caused statistically significant reduction in mean weights (A) that also translated into significant rates of mortality (B) in comparison to negative control. Data is average of 3 replicates/treatment ±SEM at p< 0.001(***); p< 0.01(**) and p< 0.05(*).
[0064] Figure 25. Figure 25 shows representative phenotypes of RFB beetles feeding on rice flour mixed with 1 pg of pure dsRNA against core MIGGS targets in RFB TcPGRP2, MspGRP2, dsCHS2, and a previously discovered target MDGP. The RFB beetles feeding on dsRNA against all MIGGS targets displayed significant mortalities at the end of 72 hours of feeding in comparison to negative control treatment (TE). The rates of mortality were significantly higher than negative control treatment and were comparable to the positive control treatment TcvATPaseE. Mortality rate was scored on a 0-3 scale were 0, 1, 2 and 3 indicated < 0, 25, 50 or > 50 mortality respectively. Data is average of 6 replicates/treatment (N=18) SEM at p< 0.001(***); p< 0.01(**) and p< 0.05(*).
[0065] Figure 26A,B. Figure 26A,B shows imageJ analyzed RT-PCR data correlating TH larval phenotypes observed when exposed to 8-16 pg pure dsRNA against a positive control (A) and a core insecticidal MIGGS RNAi target MspGRP2 (B). The transcript down regulation is correlated with the larval phenotypes observed in Figure 17.
[0066] Figure 26C,D. Figure 26C,D shows imageJ analyzed RT-PCR data correlating TH larval phenotypes observed when exposed to 8-16 pg pure dsRNA against core set of insecticidal MIGGS RNAi targets MsPGPRP2 (C) and MsCHS2 (D). The transcript down regulation is correlated with the larval phenotypes observed in Figure 17.
DETAILED DESCRIPTION Overview
[0067] RNAi-mediated gene-silencing offers a sustainable alternative approach to insect control. Most of the successful RNAi-based pest control strategies thus far employ homology dependent silencing of essential gene functions. Despite this, effective RNAi-based crop protection is lacking for Lepidopteran pests, due to their variable sensitivity to ingested double stranded RNA (dsRNA). (Terenius 0, et al. (2011). J Insect Physiol. 57(2): 231-245).
[0068] Plant pests are in constant contact with, and ingest significant amount of microbes during herbivory. (Gayatri Priya N. et al. (2012). 7(1), PLos ONE. E30768; Penuelas and Terradas (2014). 19(5): Trends Plant Sci. 278 - 280; Engel and Moran (2013). FEMS microbiol. rev. 37(5): 699-735). This interaction between ingested microbes and insect midgut is often .0 considered passive. Recent studies suggest, however, an active role of midgut specific immune responses in reducing variation of core microbial communities during insect herbivory through the activation of pattern recognition receptors (PRR). (Casanova-Torres and Goodrich-Blair (2013). Insects. 4: 320-338; Tang X, et al. (2012) 7(7) PLoS ONE:e36978; Ryu JH. et al. (2008). Science. 37(5): 777-82; Shrestha S. et al. (2009). J. Asia Pac. Entomol.12: 277-283; Buchon, N. et al. (2013). Front. Cell. Infect. Microbiol. 11: 615-626). Further, maintenance of core gut microbial communities via active immune responses and/or their containment in the midgut is key to successful herbivory.
[0069] Although, the core innate immune response pathways are conserved, their specific components are under strong selection for diversification. (Casanova-Torres and Goodrich-Blair (2013). Insects. 4: 320-338). Therefore, it is contemplated herein that these pathways provide novel and specific targets for devising sustainable pesticidal RNAi biotechnologies against insect pests. Although gut immune responses have been studied from an immunological perspective, their active manipulation via genetic engineering for pest protection is currently lacking.
[0070] Provided herein is the identification of "insect RNAi target genes" (IRTGs) involved in gut microbial clearance and/or containment induced by microbes ingested during feeding and/or active feeding (referred to herein as microbe-induced gut specific genes (MIGGS)) and examples of a novel biotechnology for insect protection via inter-specific silencing of MIGGS-IRTGs. In certain aspects, the MIGGS-IRTGs are Lepidoptera-specific. For example, in certain aspects detailed below, the insect is Manduca sexta (M sexta; Lepidoptera) (tobacco hornworm (TH)). For example, in certain aspects detailed below, the insect is Spodopterafrugiperda(fall armyworm (FAW)). For example, in certain aspects detailed below, the insect is Plutella xylostella (Diamondback moth (DBM). For example, in certain aspects detailed below, the insect is Ostrinianubilalis (European corn borer). Still, it is also considered understood that successful, feeding-induced loss of appetite, developmental defects, and/or lethality has the potential to provide protection beyond the order Lepidoptera in an orthologous manner. For example, protection against coleopteran pests such as Leptinotarsa decemlineata (Say) (Colorado potato beetle), Diabrotica spp. (Corn rootworm complex), and Tribolium castaneum (Red Flour Beetle (RFB)). Additionally, this MIGGS-RNAi technique may allow containment of disease transmitting insect vectors and/or enable further manipulation of the plant-microbe-insect interactions for devising pesticidal RNAi for crop protection. .0 [0071] In certain aspects detailed below, silencing of a target gene can result in reduced appetite and/or developmental defects and/or mortality and/or reduced fitness of the insect. In certain aspects these effects are observed after sustained feeding for at least about 24, 36, 48, or 72 hours, or any time inbetween.
Definitions
[0072] To the extent necessary to provide descriptive support, the subject matter and/or text of the appended claims is incorporated herein by reference in their entirety.
[0073] It will be understood by all readers of this written description that the exemplary embodiments described and claimed herein may be suitably practiced in the absence of any recited feature, element or step that is, or is not, specifically disclosed herein.
[0074] It is to be noted that the term "a" or "an" entity refers to one or more of that entity; for example, "a dsRNA molecule," is understood to represent one or more dsRNA molecules. As such, the terms "a" (or "an"), "one or more," and "at least one" can be used interchangeably herein.
[0075] Furthermore, "and/or" where used herein is to be taken as specific disclosure of each of the specified features or components with or without the other. Thus, the term and/or" as used in a phrase such as "A and/or B" herein is intended to include "A and B," "A or B," "A" (alone), and "B" (alone). Likewise, the term "and/or" as used in a phrase such as "A, B, and/or C" is intended to encompass each of the following embodiments: A, B, and C; A, B, or C; A or C; A or B; B or C; A and C; A and B; B and C; A (alone); B (alone); and C (alone).
[0076] It is understood that wherever aspects are described herein with the language "comprising," otherwise analogous aspects described in terms of "consisting of' and/or "consisting essentially of' are also provided.
[0077] Unless defined otherwise, technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure is related. Numeric ranges are inclusive of the numbers defining the range. Even when not explicitly identified by "and any range in between," or the like, where a list of values is recited, e.g., 1, 2, 3, or 4, unless otherwise stated, the disclosure specifically includes any range in between the values, e.g., 1to 3, 1 to 4, 2 to 4, etc.
[0078] The headings provided herein are solely for ease of reference and are not limitations of the various aspects or aspects of the disclosure, which can be had by reference to the specification as a whole.
[0079] As used herein, the term "non-naturally occurring" condition, substance, polypeptide, polynucleotide, composition, entity, plant, organism, individual, and/or any combination thereof, or any grammatical variants thereof and the like, is a conditional term that explicitly excludes, but only excludes, those forms that are well-understood by persons of ordinary skill in the art as being "naturally-occurring," or that are, or might be at any time, determined or interpreted by a judge or an administrative or judicial body to be, "naturally occurring."
[0080] As used herein, the term "identity," e.g., "percent identity" to an amino acid sequence or to a nucleotide sequence disclosed herein refers to a relationship between two or more amino acid sequences or between two or more nucleotide sequences. When a position in one sequence is occupied by the same nucleic acid base or amino acid in the corresponding position of the comparator sequence, the sequences are said to be "identical" at that position. The percentage of "sequence identity" is calculated by determining the number of positions at which the identical nucleic acid base or amino acid occurs in both sequences to yield the number of "identical" positions. The number of "identical" positions is then divided by the total number of positions in the comparison window and multiplied by 100 to yield the percentage of "sequence identity." Percentage of "sequence identity" is determined by comparing two optimally aligned sequences over a comparison window. In order to optimally align sequences for comparison, the portion of a nucleotide or amino acid sequence in the comparison window can comprise additions or deletions termed gaps while the reference sequence is kept constant. An optimal alignment is that alignment which, even with gaps, produces the greatest possible number of "identical" positions between the reference and comparator sequences. Percentage "sequence identity" between two sequences can be determined using, e.g., the program "BLAST" which is available from the National Center for Biotechnology Information, and which program incorporates the programs BLASTN (for nucleotide sequence comparison) and BLASTP (for amino acid sequence comparison), which programs are based on the algorithm of Karlin and Altschul ((1993). Proc. Natl. Acad. Sci. USA. 90(12): 5873-5877).
[0081] As used herein, the term "complementary" refers to the ability of polynucleotides to form base pairs with one another. Base pairs are typically formed by hydrogen bonds between nucleotide units in antiparallel polynucleotide strands. Complementary polynucleotide strands can base pair in the Watson-Crick manner (e.g., A to T, A to U, C to G), or in any other manner .0 that allows for the formation of duplexes. When using RNA as opposed to DNA, uracil (U) rather than thymine (T) is the base that is considered to be complementary to adenosine. However; when a U is denoted in the context of the present invention, the ability to substitute a T is implied, unless otherwise stated.
[0082] As used herein, the term "polypeptide" is intended to encompass a singular "polypeptide" as well as plural "polypeptides," and refers to a molecule composed of monomers (amino acids) linearly linked by peptide bonds (also known as amide bonds). The term "polypeptide" refers to any chain or chains of two or more amino acids, and does not refer to a specific length of the product. Thus, peptides, dipeptides, tripeptides, oligopeptides, "protein," "amino acid chain," or any other term used to refer to a chain or chains of two or more amino acids are included within the definition of "polypeptide," and unless specifically stated otherwise the term "polypeptide" can be used instead of, or interchangeably with any of these terms. The term "polypeptide" is also intended to refer to the products of post-expression modifications of the polypeptide, including without limitation glycosylation, acetylation, phosphorylation, amidation, derivatization by known protecting/blocking groups, proteolytic cleavage, or modification by non-standard amino acids. A polypeptide can be derived from a natural biological source or produced by recombinant technology, but is not necessarily translated from a designated nucleic acid sequence. Thus, it can be generated in any manner, including by chemical synthesis.
[0083] As used herein, the term "protein" refers to a single polypeptide, i.e., a single amino acid chain as defined above, but can also refer to two or more polypeptides that are associated, e.g., by disulfide bonds, hydrogen bonds, or hydrophobic interactions, to produce a multimeric protein.
[0084] As used herein, the term "nucleotide" refers to a ribonucleotide or a deoxyribonucleotide or modified form thereof, as well as an analog thereof Nucleotides include species that comprise purines, e.g., adenine, hypoxanthine, guanine, and their derivatives and analogs, as well as pyrimidines, e.g., cytosine, uracil, thymine, and their derivatives and analogs. Further, the term nucleotide also includes those species that have a detectable label, such as for example a radioactive or fluorescent moiety, or mass label attached to the nucleotide.
[0085] As used herein, the term "polynucleotide" refers to polymers of nucleotides, and includes but is not limited to DNA, RNA, DNA/RNA hybrids including polynucleotide chains of regularly and/or irregularly alternating deoxyribosyl moieties and ribosyl moieties (i.e., wherein .0 alternate nucleotide units have an -OH, then and -H, then an -OH, then an -H, and so on at the 2'position of a sugar moiety), and modifications of these kinds of polynucleotides, wherein the attachment of various entities or moieties to the nucleotide units at any position are included. The term "polynucleotide" is also intended to encompass a singular nucleic acid as well as plural nucleic acids, and refers to an isolated nucleic acid molecule or construct, e.g., messenger RNA (mRNA) or plasmid DNA (pDNA). A polynucleotide can comprise a conventional phosphodiester bond or a non-conventional bond (e.g., an amide bond, such as found in peptide nucleic acids (PNA)). A polynucleotide can be single stranded or double stranded.
[0086] As used herein, the term "nucleic acid" refers to any one or more nucleic acid segments, e.g., DNA or RNA fragments, present in a polynucleotide. By "isolated" nucleic acid or polynucleotide is intended a nucleic acid molecule, DNA or RNA, which has been removed from its native environment. For example, a recombinant polynucleotide encoding a polypeptide subunit contained in a vector is considered isolated as disclosed herein. Further examples of an isolated polynucleotide include recombinant polynucleotides maintained in heterologous host cells or purified (partially or substantially) polynucleotides in solution. Isolated RNA molecules include in vivo or in vitro RNA transcripts of polynucleotides. Isolated polynucleotides or nucleic acids further include such molecules produced synthetically. In addition, polynucleotide or a nucleic acid can be or can include a regulatory element such as a promoter, ribosome binding site, or a transcription terminator.
[0087] As used herein, a "coding region" is a portion of nucleic acid comprising codons translatable into amino acids. Although a "stop codon" (TAG, TGA, or TAA) is not translated into an amino acid, it can be considered to be part of a coding region, but any flanking sequences, for example 5' untranslated regions (5' UTRs; also known as a leader sequence), 3' untranslated regions (3' UTRs; also known as a trailer sequence), promoters, ribosome binding sites, transcriptional terminators, introns, and the like, are not part of a coding region. Two or more coding regions can be present in a single polynucleotide construct, e.g., on a single vector, or in separate polynucleotide constructs, e.g., on separate (different) vectors. Furthermore, any vector can contain a single coding region, or can comprise two or more coding regions, e.g., a single vector can separately encode a selection marker gene and a gene of interest. In addition, a vector, polynucleotide, or nucleic acid can encode heterologous coding regions, either fused or unfused to a nucleic acid encoding a polypeptide subunit or fusion protein as provided herein. Heterologous coding regions include without limitation specialized elements or motifs, such as a secretory signal peptide or a heterologous functional domain.
[0088] A variety of transcription regulatory regions are known to those skilled in the art. These include, without limitation, transcription regulatory regions that function in vertebrate cells, such as, but not limited to, promoter and enhancer segments from cytomegaloviruses (the immediate early promoter, in conjunction with intron-A), simian virus 40 (the early promoter), and retroviruses (such as Rous sarcoma virus). Other transcription regulatory regions include those derived from vertebrate genes such as actin, heat shock protein, bovine growth hormone and rabbit p-globin, as well as other sequences capable of controlling gene expression in eukaryotic cells. Additional suitable transcription regulatory regions include tissue-specific promoters and enhancers.
[0089] Similarly, a variety of translation regulatory elements are known to those of ordinary skill in the art. These include, but are not limited to ribosome binding sites, translation initiation and termination codons, and elements derived from picornaviruses (particularly an internal ribosome entry site, or RES).
[0090] As used herein, the term "vector" is nucleic acid molecule as introduced into a host cell or organelle, thereby producing a transformed host cell or organelle. A vector can include nucleic acid sequences that permit it to replicate in a host cell, such as an origin of replication. A vector can also include one or more selectable marker gene and other genetic elements known in the art. Illustrative types of vectors include plasmids, phages, viruses and retroviruses.
[0091] As used herein, the term "transformed" cell or organelle, or a "host" cell organelle, is a cell or organelle into which a nucleic acid molecule has been introduced by molecular biology techniques. As used herein, the term transformation encompasses those techniques by which a nucleic acid molecule can be introduced into such a cell or organelle, including transfection with viral vectors, transformation with plasmid vectors, and introduction of naked DNA by electroporation, lipofection, and particle gun acceleration. A transformed cell or a host cell can be a bacterial cell or a eukaryotic cell.
[0092] As used herein, the term "expression" refers to a process by which a gene produces a biochemical, for example, a polynucleotide or a polypeptide. The process includes any manifestation of the functional presence of the gene within the cell including, without limitation, gene knockdown as well as both transient expression and stable expression. It includes .0 without limitation transcription of the gene into messenger RNA (mRNA), and the translation of such mRNA into polypeptide(s). It also includes without limitation transcription of the gene into an RNA molecule that is not translated into a polypeptide but is capable of being processed by cellular RNAi mechanisms. If the final desired product is a biochemical, expression includes the creation of that biochemical and any precursors. Expression of a gene produces a "gene product." As used herein, a gene product can be either a nucleic acid, e.g., an RNA produced by transcription of a gene or a polypeptide that is translated from a mRNA transcript. Gene products described herein further include nucleic acids with post transcriptional modifications, e.g., polyadenylation, or polypeptides with post translational modifications, e.g., methylation, glycosylation, the addition of lipids, association with other protein subunits, proteolytic cleavage, and the like.
[0093] As used herein the term "engineered" includes manipulation of nucleic acid or polypeptide molecules by synthetic means (e.g. by recombinant techniques, in vitro peptide synthesis, by enzymatic or chemical coupling of peptides or some combination of these techniques).
[0094] As used herein, the term "hpRNA" refers to hairpin RNA comprising a single stranded loop region and a base-paired stem of an inversely repeated sequence. hpRNA can be generated from an hpRNA construct (or vector) and/or an hpRNA transgene comprising an inversely-repeated sequence of the RNAi target gene with a spacer region between the repeats. The RNA transcribed from such a sequence self-hybridizes to form a hairpin structure. The stem can be used as a substrate for the generation of siRNAs, but few or none are generated from the loop. Since a spacer region is needed for the stability of the transgene construct, but is not involved in siRNA production, an intron sequence is often used in this position. (Watson JM, et al. (2005). FEBS Letters. 579: 5982-8987).
[0095] As used herein, the term "siRNA" refers to small (or short) interfering RNA (or alternatively, silencing RNA) duplexes that are capable of inducing the RNA interference (RNAi) pathway. These molecules can vary in length (generally between 18-30 base pairs) and contain varying degrees of complementarity to their target mRNA in the antisense strand. Some, but not all, siRNA have unpaired overhanging bases on the 5' or 3' end of the sense strand and/or the antisense strand. The term "siRNA" includes duplexes of two separate strands, as well as single strands that can form hairpin structures comprising a duplex region.
[0096] As used herein, the phrase "duplex region" refers to the region in two complementary or substantially complementary polynucleotides that form base pairs with one another, either by Watson-Crick base pairing or any other manner that allows for a stabilized duplex between polynucleotide strands that are complementary or substantially complementary. For example, a polynucleotide strand having 21 nucleotide units can base pair with another polynucleotide of 21 nucleotide units, yet only 19 bases on each strand are complementary or substantially complementary, such that the "duplex region" has 19 base pairs. The remaining bases may, for example, exist as 5' and 3' overhangs. Further, within the duplex region, 100% complementarity is not required; substantial complementarity is allowable within a duplex region. Substantial complementarity as used herein refers to 79% or greater complementarity. For example, a mismatch in a duplex region consisting of 19 base pairs results in 94.7% complementarity, rendering the duplex region substantially complementary.
[0097] As used herein, the phrase "gene silencing" refers to a process by which the expression of a specific gene product is lessened or attenuated. Silencing of a gene does not require that the expression or presence of the gene product is completely absent, but that in the context (e.g., comparing expression of a target gene in a plant expressing a gene silencing nucleic acid compared to a control plant or the health of an insect feeding on a gene silencing nucleic acid compared to a control insect), an observable effect in comparison to a control is observed. While gene silencing can take place by a variety of pathways, unless specified otherwise, as used herein, gene silencing refers to decreases in gene product expression that results from RNA interference (RNAi) as understood by one of ordinary skill in the art. The level of gene silencing can be measured by a variety of means, including, but not limited to, measurement of transcript levels by Reverse transcription polymerase chain reaction (PCR), Northern Blot Analysis, B DNA techniques, transcription-sensitive reporter constructs, expression profiling (e.g. DNA chips), and related technologies. Alternatively, the level of silencing can be measured by assessing the level of the protein encoded by a specific gene. This can be accomplished by performing a number of studies including Western Analysis, measuring the levels of expression of a reporter protein that has e.g. fluorescent properties (e.g. GFP) or enzymatic activity (e.g. alkaline phosphatases), or several other well-known procedures. Further, gene silencing can be assessed by its effect on a pest insect such as resulting in reduced appetite and/or developmental defects and/or mortality of an insect.
[0098] As used herein, the term "control" is consistent with its well-established scientific use that refers to a standard of comparison recognized by one of ordinary skill in the art as having .0 a representative level of expression, phenotype, resistance, feeding, mortality, development, etc. Further, one of ordinary skill in the art will recognize, for example, that a statistical outlier and/or non-representative result produced by chance, abnormal environmental condition, manipulation, or other reason, that varies from a standard representation, would not be an appropriate control.
[0099] As used herein, "microbe-induced gut specific genes (MIGGS)" refers to a gene or group of genes expressed in the insect midgut in response to microbes ingested during normal process of insect feeding and primarily functioning to clear or respond to the ingested microbes and/ or contain the microbes to insect gut via maintenance of midgut structural integrity.
[0100] As used herein, "actively feeding stage of the insect" refers to all feeding stages of insects with both complete and incomplete metamorphosis.
Nucleic Acids for Gene Silencing
[0101] Provided herein are nucleic acid molecules for use in, among other things, crop protection from insect pests. In certain aspects disclosed herein, the nucleic acid molecules are isolated. The nucleic acid molecules specifically target certain insect genes (referred to herein interchangeably as "target genes," "RNAi target genes," "insect RNAi target genes," and "IRTGs"), in insects for gene silencing. For example, in certain aspects, the nucleic acid molecules target certain insect microbe-induced gut gene (MIGGS) RNAi targets. In certain aspects, the silencing of a target gene occurs when a nucleic acid molecule of this disclosure is ingested by an insect. In certain aspects, the target gene is an insect gene that is implicated in insect immune responses (type 1 MIGGS RNAi target). A critical immune response gene is a genetically tractable nuclear or cytoplasmic loci that is important for providing cellular and/or humoral defense in insects against internal microorganisms, external microorganisms, and/or other insect parasites. In certain aspects, the immune response genes (type 1 MIGGS RNAi target) can also be a pattern recognition receptor (PRR) gene (Casanova-Torres and Goodrich Blair (2013). Insects. 4:320-338). A PPR gene is a genetically tractable loci of an insect that encodes soluble or membrane bound proteins that recognize signatures associated with and/or released by microorganisms. PRR genes can activate or be activated by the immune response pathways to minimize microbial infection and can be co-regulated by the immune deficiency (ID) pathway (Tang X, et al. (2012) 7(7) PLoS ONE:e36978; Ryu JH. et al. (2008). Science. 37(5): 777-82; Shrestha S. et al. (2009). In certain aspects the PRR type genes are co-regulated by the immune deficiency (IMD) pathway in TH were identified, these genes having been recently summarized. (Casanova-Torres and Goodrich-Blair (2013). Insects (4): 320-338; Zhong X, et al. (2012). Insect Biochem. Mol. Biol. 42(7): 514-524); Zhang X, et al. (2015). Insect Biochem. Mol. Biol. 62:38-50; Cao X, et al. (2015). Insect Biochem.Mol. Biol. 62:64-74; Kanost MR, et al. (2016). Insect Biochem. Mol. Biol 76:118-147). In certain aspects, the target gene is an insect gene that is necessary for structural integrity of insect organs including the mid-gut and also facilitates the containment of the ingested microbes to the insect gut. (type 2 MIGGS RNAi target). In certain aspects, the target gene is an insect midgut structural component gene (type 2) (Odman-Naresh et al. (2013). PLoS ONE 8:e82015. 10.1371/journal.pone.0082015). A midgut structural component gene is a genetically tractable loci in an insect that encodes chitin fibrils, proteins, or glycoproteins that form a protective sac-like structure called peritrophic matrix enveloping the insect food bolus/midgut also functioning to contain the ingested microbes in the gut (Engel and Moran (2013). FEMS MicrobiolRev. 37 699-735). In certain aspects, the target genes (type 1 and 2 MIGGS RNAi targets) are predominantly expressed in the insect midgut, for example, abundantly and/or exclusively expressed in the larval and/or adult insect midgut in response to active feeding and/or microbial infection and/or responding to microbes ingested during feeding. In certain aspects, the target gene is induced predominantly in a midgut specific manner during active feeding (type 1 and type 2 MIGGS RNAi targets). The midgut abundance of both type 1 and 2 MIGGS RNAi target genes may mitigate problems associated with reduced amounts of bioavailability.
[0102] Representative examples of insect MIGGS RNAi target genes and their nucleic acid sequences identified from published literature are provided herein. In certain aspects, the target gene is one or more ofM sexta-Hemolin (MsHEM), M. sexta-Serine proteinase homolog 3 (MsSPH-3), M. sexta-Peptidoglycan recognition protein 2 (MsPGRP2), M sexta-Beta-1, 3 glycan-recognition protein 2 (MspGRP2), M. sexta-Relish family protein 2A (MsREL2A), M sexta-Dorsal (MsDor), M sexta-Spatzle (MsSPZ1A), M sexta-Toll receptor (MsTOLL), M.sexta-Scolexin A (MsSCA1), M. sexta-Hemolymph proteinase 18 (MsHP18), M sexta Transferrin (MsTRN), M. sexta-Arylphorin beta subunit (MsARP), M sexta-Chymotrypsinogen like protein 1 (MsCTL1), M sexta-Valine Rich Midgut Protein (MsVMP1), M. sexta-Imd (MsImd), M sexta-FADD (MsFADD), M. sexta-Dredd (MsDRD), M. sexta-Relish F (MsReIF), M. sexta-Cdc42 (MsCdc42), M sexta-Dsorl (MsDsorl), M sexta-Fos (MsFos), M sexta-Jra (MsJra), M. sexta-Caudal (MsCAD), M sexta-Atg8 (MsAtg8), M sexta-Atgl3 (MsAtgl3), M sexta-IAP1(MsIAP1), M. sexta-Chitin synthase 2 (MsChs2), M. sexta- Beta fructofuranosidase 1 (MsSucl), and orthologs thereof
[0103] In certain aspects, the target gene is one or more ofM. sexta-Hemolin (MsHEM), M. sexta-Serine proteinase homolog 3 (MsSPH-3), M sexta-Peptidoglycan recognition protein 2 (MsPGRP2), M. sexta-Beta-1, 3-glycan-recognition protein 2 (MspGRP2), M sexta-Relish family protein 2A (MsREL2A), M. sexta-Dorsal (MsDor), M. sexta-Sptzle (MsSPZ1A), M. sexta-Toll receptor (MsTOLL), Msexta-Scolexin A (MsSCA1), M sexta-Hemolymph proteinase 18 (MsHP18), M. sexta-Transferrin (MsTRN), M. sexta-Arylphorin beta subunit (MsARP), M sexta-Chymotrypsinogen-like protein 1 (MsCTL1), M sexta-Valine Rich Midgut Protein (MsVMIP1), M sexta-Imd (Mslmd), M. sexta-FADD (MsFADD), M sexta-Dredd (MsDRD), M sexta-Relish F (MsReIF), M sexta-Cdc42 (MsCdc42), M. sexta-Dsorl (MsDsorl), M sexta-Fos (MsFos), M sexta-Jra(MsJra), M. sexta-Caudal (MsCAD1), M sexta-Atg8 (MsAtg8), M sexta
Atg13 (MsAtg13), M. sexta-IAP1(MsIAP1), M sexta-Chitin synthase 2 (MsChs2), and M sexta Beta fructofuranosidase 1 (MsSucl), , M. sexta-Sickie (MsSck), M. sexta-Akirin (MsAki), M sexta-Cactus (MsCac),M. sexta- Gloverin (MsGlv) andM sexta-Beta--tubulin (MsTub).
[0104] In certain aspects, the target gene is an ortholog of one or more of M sexta Hemolin (MsHEM), M. sexta-Serine proteinase homolog 3 (MsSPH-3), M. sexta-Peptidoglycan recognition protein 2 (MsPGRP2), M sexta-Beta-1, 3-glycan-recognition protein 2 (MspGRP2), M. sexta-Relish family protein 2A (MsREL2A), M sexta-Dorsal (MsDor), M. sexta-Sptzle (MsSPZ1A), M sexta-Toll receptor (MsTOLL), M.sexta-Scolexin A (MsSCA1), M. sexta Hemolymph proteinase 18 (MsHP18), M. sexta-Transferrin (MsTRN), M. sexta-Arylphorin beta subunit (MsARP), M sexta-Chymotrypsinogen-like protein 1 (MsCTL1), M. sexta-Valine Rich Midgut Protein (MsVMP), M. sexta-Imd (MsImd), M sexta-FADD (MsFADD), M. sexta Dredd (MsDRD), M sexta-Relish F (MsReIF), M. sexta-Cdc42 (MsCdc42), M. sexta-Dsorl (MsDsorl), M sexta-Fos (MsFos), M. sexta-Jra (MsJra), M. sexta-Caudal (MsCAD1), M sexta Atg8 (MsAtg8), M sexta-Atgl3 (MsAtgl3), M. sexta-IAP1(MsIAP1), M. sexta-Chitin synthase 2 (MsChs2), M sexta- Beta fructofuranosidase 1 (MsSucl), and other IMD pathway or structural integrity genes.
[0105] One of ordinary skill in the art would understand that nucleic acid molecules can be, for example, deoxyribonucleic acid (DNA) or ribonucleic acid (RNA). In certain aspects of any target gene silencing nucleic acid molecule described anywhere herein, the nucleic acid .0 molecule is a DNA molecule. In certain aspects of any target gene silencing nucleic acid molecule described anywhere herein, the nucleic acid molecule is a RNA molecule. In certain aspects of any target gene silencing nucleic acid molecule described anywhere herein, the RNA molecule is a double stranded molecule (dsRNA), for example, for use in the RNA interference (RNAi) process. As used herein, a dsRNA molecule is a RNA molecule comprising at least one annealed, double stranded region. In certain aspects, the double stranded region comprises two separate RNA strands annealed together. In certain aspects, the double stranded region comprises one RNA strand annealed to itself, for example, as can be formed when a single RNA strand contains an inversely repeated sequences with a spacer in between. One of ordinary skill in the art will understand that complementary nucleic acid sequences are able to anneal to each other but that two sequences need not be 100% complementary to anneal. The amount of complementarity needed for annealing can be influenced by the annealing conditions such as temperature, pH, and ionic condition. In certain aspects, the annealed RNA sequences are 100% complementary across the annealed region. In certain aspects, the annealed RNA sequences are less than 100% complementary across the annealed region but have enough complementarity to anneal within their environment, such as in a host cell or the gut of an insect. In certain aspects, the annealed RNA sequences are substantial complementarity as defined elsewhere herein.
[0106] It is contemplated that the nucleic acid molecules disclosed anywhere herein for the silencing of target genes derive their specificity from comprising a nucleic acid sequence that is complementary or substantially complementary to at least a portion of a target gene sequence. Substantially complementary sequences, however, may be more likely to have reduced specificity and produce off-target effects. As referred to anywhere herein, a target gene sequence can include at least the target gene protein coding region, the 5' untranslated region (5' UTR), and/or the 3' untranslated region (3' UTR) and any portion or combination thereof For example, predicted UTR regions can be identified using previously established criteria (Siepel, et al. (2005). Genome Res. 15: 1034-1050) when corresponding genomic sequences are available.
[0107] In certain aspects, an isolated double stranded RNA (dsRNA) molecule comprises a nucleic acid sequence complementary to about 21 to 2000 contiguous nucleotides of a target gene sequence discloses anywhere herein. For example, in certain aspects, an isolated double stranded RNA (dsRNA) molecule comprises a nucleic acid sequence complementary to about any of21,22,23,24,25, 30, 40, 50, 60, 100, 120,200,240, 300, 400, 500, 600, 650, 750, 1000 to about any of23, 24, 25, 30, 40, 50, 100, 200, 300, 400, 500, 600, 650, 750, 1000, or 2000 contiguous nucleotides of a target gene sequence. For example, in certain aspects, an isolated dsRNA molecule comprises a nucleic acid sequence complementary to about 100 to 1000 or about 200 to 1000 contiguous nucleotides of a target gene sequence. For example, in certain aspects, an isolated dsRNA molecule comprises a nucleic acid sequence complementary to about 100 to 1000 or about 200 to 1000 contiguous nucleotides of the protein coding region of a target gene sequence. For example, in certain aspects, an isolated dsRNA molecule comprises a nucleic acid sequence complementary to about 100 to 1000 or about 200 to 1000 contiguous nucleotides of the 5' UTR region or the 3' UTR region of a target gene sequence. In certain aspects, the isolated dsRNA molecule comprises a nucleic acid sequence complementary to a contiguous region comprising at least about 50%, 60%, 70%, 80%, 90%, 95%, 9 8 %, 99%, or 100% of the length of the target gene sequence protein coding region, the target gene sequence 5' UTR region, the target gene 3' UTR region, and/or any combination thereof For example, if a target gene sequence protein coding region is determined to be 200 nucleotides long, then an isolated dsRNA molecule comprising a nucleic acid sequence complementary to a contiguous region comprising 95% of the length of the target gene sequence protein coding region would be complementary to a contiguous region 190 nucleotides long.
[0108] In certain aspects of any target gene silencing nucleic acid molecule described anywhere herein, including dsRNA molecules for RNAi, the target gene comprises one or more of the nucleic acid sequence of SEQ ID NO: 1-14, 16-29, 31-69, 70-75, 76-88, 89-105, and 106 110. In certain aspects, the target gene comprises the nucleic acid sequence of SEQ ID NO: 3 or 14. Thus, in certain aspects, the isolated dsRNA molecule comprises a nucleic acid sequence complementary to about any of 21, 22, 23, 24, 25, 30, 40, 50, 100, 200, 300, 400, 500, 600, 650, 750,1000to aboutany of23,24,25,30,40, 50,100,200,300,400,500,600,650,750, 1000,or 2000 contiguous nucleotides of a target gene sequence comprising a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 1-14, 16-29, 31-69, 70-75, 76-88, 89-105, and 106-110. For example, in certain aspects, the isolated dsRNA molecule comprises a nucleic acid sequence complementary to about 100 to 1000 or about 200 to 1000 contiguous nucleotides of a target gene sequence comprising a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 1-14, 16-29, 31-69, 70-75, 76-88, 89-105, and 106-110. In certain aspects, the isolated dsRNA comprises a nucleic acid sequence complementary to about 200 to 1000 contiguous nucleotides of the protein coding region of a target gene sequence comprising a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 1-14, 16-29, 31-69, 70-75, 76-88, 89-105, and 106-110. In certain aspects, the isolated dsRNA molecule comprises a nucleic acid sequence complementary to a contiguous region comprising at least about 50%, 60%, 70%, 80%, 90%, 95%, 9 8 %, 9 9 %, or 100% of the length of the target gene sequence protein coding region, the target gene 5' UTR region, and/or the target gene 3' UTR region. In certain aspects, the isolated dsRNA molecule comprises a nucleic acid sequence complementary to a contiguous region comprising at least about 50%, 60%, 70%, 80%, 90%, 95%, 9 8 %, 9 9 %, or 100% of the length of a sequence selected from the group consisting of SEQ ID NOs: 1-14, 16 29,31-69,70-75,76-88, 89-105, and 106-110.
[0109] In certain aspects of any target gene silencing nucleic acid molecule described anywhere herein, the nucleic acid molecule can form siRNA. Thus, certain aspects provide for an siRNA molecule derived from the processing of a dsRNA molecule for silencing a target gene disclosed herein.
Insecticidal Compositions
[0110] Certain aspects of the disclosure provide for an insecticidal composition comprising a nucleic acid molecule disclosed anywhere herein for silencing a target gene, including long dsRNA, hpRNA, and siRNA. In certain aspects, the insecticidal composition also comprises a synthetic carrier or a microbial conduit. For example, a microbial conduit can be a microorganism that has a natural capacity or is engineered to produce and/ or deliver dsRNA to increase its bioavailability and/or biostability for causing RNA interference. Representative examples include plant growth promoting organisms, normal commensal and/or symbiotic microorganisms associated with the target insect pest or pest target host or host cultivation range etc. from an insect engineered or identified from natural populations to produce and/or deliver dsRNA. In certain aspects a microbial conduit can be used as a direct topical application on a whole plant or coated onto a seed or mixed with growth media or transmitted through fertilizer or irrigation, etc. In certain aspects, the nucleic acid molecule of the insecticidal composition is conjugated to the synthetic carrier. For example, a synthetic carrier can be an inert chemical compound with a natural or engineered affinity to bind (conjugate) a dsRNA molecule to increase its biostability and/or bioavailability for causing RNA interference. In certain aspects, a synthetic carrier comprises a combination of inert chemicals or nanoparticles that upon combining and/or individually have a net positive charge or general affinity to bind to negatively charged dsRNA. Representative examples include chitosan, liposomes, carbon quantum dots, .0 biodegradable particles of plant (e.g. coconut coir or grain flour, etc.) or soil (e.g. calcified clay) origin etc. In certain aspects, the dsRNA conjugated with a synthetic carrier can be used as a direct topical application directly and/or after aerosolization on a whole plant or coated onto a seed or mixed with growth media or transmitted through fertilizer or irrigation, etc. In certain aspects, dsRNA or a composition comprising dsRNA can be used as a direct topical spray on application to whole plant, coated onto a seed or mixed with growth media or transmitted through fertilizer or irrigation or combined with plant growth promoting microbes etc.
Recombinant Constructs
[0111] Certain aspects of this disclosure provide for a recombinant nucleic acid construct, such as a DNA vector, comprising and/or encoding a nucleic acid molecule disclosed anywhere herein for silencing a target gene, including long dsRNA, hpRNA, and siRNA. Certain aspects provide for recombinant nucleic acid constructs comprising and/or encoding an RNAi precursor of a nucleic acid molecule disclosed anywhere herein for silencing a target gene, including long dsRNA, hpRNA, and siRNA.
[0112] Certain aspects of this disclosure provide for a recombinant nucleic acid construct, such as a DNA vector, comprising a target gene silencing sequence for silencing a target gene described anywhere herein. In certain aspects, a recombinant DNA construct comprises a gene silencing sequence comprising about any of 21, 22, 23, 24, 25, 30, 40, 50, 60, 100, 120, 200, 240, 300, 400, 500, 600, 650, 750, 1000 to about any of23, 24, 25, 30, 40, 50, 100, 200, 300, 400, 500, 600, 650, 750, 1000, or 2000 contiguous nucleotides of a target gene sequence disclosed anywhere herein. In certain aspects, a recombinant DNA construct comprises a gene silencing sequence comprising about 100 to 1000 or about 200 to 1000 contiguous nucleotides of a target gene sequence. In certain aspects, the gene silencing sequence comprises about 100 to 1000 or about 200 to 1000 contiguous nucleotides of the protein coding region of the target gene sequence. In certain aspects, the gene silencing sequence comprises about 100 to 1000 or about 200 to 1000 contiguous nucleotides of the 5' UTR region or the 3' UTR region of the target gene sequence. In certain aspects, the gene silencing sequence comprises at least 50%, 60%, 70%, 80% , 90%, 95%, 9 8 %, 9 9 %, or 100% contiguously of the length of target gene sequence protein coding region, the target gene sequence 5' UTR region, target gene sequence 3' UTR region and/or any combination thereof
[0113] In certain aspects, the target gene comprises a nucleic acid sequence selected from .0 the group consisting of SEQ ID NOs: 1-14, 16-29, 31-69, 70-75, 76-88, 89-105, and 106-110. In certain aspects, the gene silencing sequence comprises about 100 to 1000 or about 200 to 1000 contiguous nucleotides of a sequence selected from the group consisting of SEQ ID NOs: 1-14, 16-29, 31-69, 70-75, 76-88, 89-105, and 106-110. In certain aspects, the gene silencing sequence comprises at least 50%, 60%, 70%, 80%, 90%, 95%, 98%, 99%, or 100% contiguously of the length of a sequence selected from the group consisting of SEQ ID NOs: 1-14, 16-29, 31-69, 70 75,76-88, 89-105, and 106-110.
[0114] In certain aspects, the recombinant DNA construct has a gene silencing sequence operably linked to one or more promoters for the expression of a dsRNA molecule that silences the target gene when ingested by an insect. Thus, in certain aspects, the construct is an expression vector. Representative promoters for use in expressing a dsRNA molecule include, but are not limited to, CaMV35S or ZmUbil promoters etc. In certain aspects, the expression vector can target single or multiple insect RNAi target genes, for example, the vector could comprise one or more gene silencing sequences or could employ multiple vectors to target multiple insect RNAi target genes or chimeric dsRNA molecules.
Host Cells and Plants
[0115] Provided herein are host cells, plants, and plants parts comprising, expressing, processing, and the like a dsRNA as described anywhere herein for inducing RNAi in an insect. In certain aspects, a host cell comprises a dsRNA molecule, siRNA molecule, a polynucleotide encoding a dsRNA molecule, and/or a construct or a dsRNA encoding segment thereof described anywhere herein. Representative examples of host cells include bacterial cells, fungal cells, yeast cells, plant cells, plant organelles (e.g., including plastids), and mammalian cells. In certain aspects, the host cell is a bacterial or plant cell. In certain aspects, the host cell is a transgenic and/or transplastomic plant cell. One of ordinary skill will understand that there are many well known methods for introducing a nucleic acid, such as a vector, into a host cell including well known methods for generating transgenic and/or transplastomic plant cells. In certain aspects, the hose cell expresses a dsRNA molecules and/or produces siRNA to silence a target gene. In certain aspects, a transgenic and/or transplastomic plant can comprise a dsRNA molecule, siRNA, a polynucleotide encoding a dsRNA, and/or a construct or a dsRNA encoding segment thereof. In certain aspects, at least one cell of a transgenic and/or transplastomic plant expresses a dsRNA molecule and/or produces a siRNA for silencing a target gene. Certain aspects provide for a seed, part, tissue, cell, or organelle of a plant described herein, wherein said seed, part, tissue, cell, or organelle comprises a dsRNA molecule and/or the siRNA for silencing a target gene.
Methods of Insect Control
[0116] Also provided for herein are various methods of using a dsRNA molecule or vector encoding such dsRNA described anywhere herein for inducing RNAi in an insect and/or silencing a target gene. In certain aspects, this provides for control of insect pests.
[0117] Certain aspects provide for a method of silencing an insect immune response gene and/or an insect gene encoding for structural components of the insect midgut. In certain aspects, a method provides for the silencing an insect MIGGS-IRTG. Such method comprises providing for ingestion through spray, drenches, granules, seed coating or plant-incorporated protectant, or the like, to an insect an isolated dsRNA (pure or crude extract), siRNA, insecticidal composition, host cell, transgenic and/or transplastomic plant, and/or the seed, part, tissue, cell, or organelle thereof described anywhere herein.
[0118] Certain aspects provide for a method of silencing an insect immune response gene and/or an insect gene encoding for structural components of the insect midgut. In certain aspects, a method provides for the silencing an insect MIGGS-IRTG. Such method comprises providing for ingestion through spray, drenches, granules, seed coating or plant-incorporated protectant, or the like, to an insect an isolated dsRNA, siRNA, insecticidal composition, host cell, transgenic and/or transplastomic plant, and/or the seed, part, tissue, cell, or organelle thereof described anywhere herein.
[0119] Certain aspects provide for protecting a plant, such as a crop plant, from an insect pest including but not limited to pests of the order Lepidoptera like Manduca sexta (tobacco hornworm), Spodoptera frugiperda (fall armyworm), Ostrinia nubilalis (European corn borer), Plutella xylostella (Diamondback moth) or pests of the order Coleoptera like Leptinotarsa decemlineata Say (Colorado potato beetle), Diabrotica spp. (Corn rootworm complex), Tribolium castaneum (Red flour beetle), Popil/ajaponica(Japanese beetle), Agrilusplanipennis (Emerald ash borer) or pests of the order Hemiptera like Diaphorinacitri (Asian citrus psyllid), Cimex lectularius (Bed bug) or pests of the order Blattodea like all species of cockroaches and termites or insect pests of the order Diptera like all species of Mosquitoes and flies etc. Representative examples of plant hosts include, but are not restricted to, Zea mays L (corn), .0 Sorghum bicolor (sorghum), Setaria italica (fox tail millet), Pennisetum glaucum (Pearl millet), Solanum tuberosum (potato), Oryza sativa (rice), Lycopersicon esculentum (tomato), Solanum melongena (eggplant), all cultivars of Brassicaoleracea family, Citrus sinensis (Orange), trees of Oleaceae family and crops of Rosaceae etc. Such methods comprise, for example, topically applying to the plant the isolated dsRNA (pure or crude extract), the siRNA, and/or the insecticidal composition described anywhere herein, and providing the plant in the diet of the insect pest. In certain aspects the dsRNA molecule is topically applied by expressing the dsRNA in a microbe followed by topically applying the microbe onto the plant and/or seed.
[0120] Certain aspects provide for producing a plant resistance to a pest insect of said plant. Such methods comprises transforming the plant with a polynucleotide encoding a dsRNA and/or a construct or a dsRNA encoding segment describe anywhere herein, wherein the plant expresses a dsRNA and/or siRNA and/or the plant comprises a dsRNA and/or siRNA containing insecticidal compositions described anywhere herein, for silencing a target gene. In certain aspects, the transformed plant is more resistant to a pest insect of said plant than untransformed plants.
[0121] Certain aspects provide for improving crop yield. Such methods comprise growing a population of crop plants transformed with a polynucleotide encoding a dsRNA and/or the construct or a dsRNA encoding segment thereof described anywhere herein, wherein the plant expresses a dsRNA and/or siRNA and/or the plant comprises a dsRNA and/or siRNA containing insecticidal compositions described anywhere herein, for silencing a target gene. In certain aspects, a population of transformed plants produces higher yields in the presence of pest insect infestation than a control population of untransformed plants.
[0122] Certain aspects of the disclosure provide for an insecticidal composition comprising a nucleic acid molecule disclosed anywhere herein for silencing a target gene, including long dsRNA, hpRNA, and siRNA. In certain aspects, the insecticidal composition also comprises a synthetic carrier or a microbial conduit. For example, a microbial conduit can be a microorganism that has a natural capacity or is engineered to produce and/ or deliver dsRNA to increase its bioavailability and/or biostability for causing RNA interference. Representative examples include plant growth promoting organisms, normal commensal and/or symbiotic microorganisms associated with the target insect pest or parasites and/or natural enemies of the target pest or pest target host or host cultivation range etc. from an insect or parasite and/or natural enemies of the target pest engineered or identified from natural populations containing microbial conduit to produce and/or deliver dsRNA and/or drive the transmission of such microbial conduits into natural populations of insect pests as a control option. In certain aspects a microbial conduit can be used as a direct topical application on a whole plant or coated onto a seed or mixed with growth media or transmitted through fertilizer or irrigation, etc. In certain aspects, the nucleic acid molecule of the insecticidal composition is conjugated to the synthetic carrier. For example, a synthetic carrier can be an inert chemical compound with a natural or engineered affinity to bind (conjugate) a dsRNA molecule to increase its biostability and/or bioavailability for causing RNA interference. In certain aspects, a synthetic carrier comprises a combination of inert chemicals or nanoparticles that upon combining and/or individually have a net positive charge or general affinity to bind to negatively charged dsRNA. Representative examples include chitosan, liposomes, carbon quantum dots, biodegradable particles of plant (e.g. coconut coir or grain flour, etc.) or soil (e.g. calcified clay) origin etc. In certain aspects, the dsRNA conjugated with a synthetic carrier can be used as a direct topical application directly and/or after aerosolization on a whole plant or coated onto a seed or mixed with growth media or transmitted through fertilizer or irrigation, etc. In certain aspects dsRNA or composition comprising the dsRNA can be used as a direct topical spray on application to whole plant, coated onto a seed or mixed with growth media or transmitted through fertilizer or irrigation or combined with plant growth promoting microbes etc.
[0123] Certain aspects provide for producing a plant resistant against a pest insect of said plant. Such methods comprise first transforming a plant cell with a polynucleotide encoding the dsRNA and/or the construct or a dsRNA encoding segment described anywhere herein. Next, a plant is regenerated from the transformed plant cell. The plant is then grown under conditions suitable for the expression of the dsRNA. In certain aspects, the transformed plant confers genetically tractable (maternal and/or paternal inherited) gain of function phenotypically manifested as an ability to impair the normal feeding and/or growth and/or development and/or reproductive success of the target plant pest and is consequently resistant to the plant pest insect compared to a control untransformed plant.
[0124] In certain aspects of any of the aforementioned methods, the insect larvae ingest the dsRNA. In certain aspects of any of the aforementioned methods, ingestion of the dsRNA induces a melanotic response in the insect larvae. In certain aspects of any of the aforementioned methods, ingestion of the dsRNA results in perturbation of gut microbial homeostasis. In certain aspects of any of the aforementioned methods, ingestion of the dsRNA results in defective clearance of opportunistic microbes. In certain aspects of any of the aforementioned methods, ingestion of the dsRNA results in defective containment of gut microbes.
[0125] One of ordinary skill in the art will recognize that, the inventors have demonstrated midgut specific expression of a representative number of MIGGS-IRTGS in TH in response to their feeding on a lab strain of Escherichia coli (E. coi) bacteria. For example, a set of 20 MIGGS-IRTGS (SEQ ID NOs: 1-9, 11, 14, 31, 39, 43, 44, 71-75) that are induced in TH larvae feeding on an induction medium (Wang et al. (2006). J. Biol. Chem. 281(14): 9271-9278) is disclosed herein. It was previously reported that these twenty genes are expressed abundantly in the gut (Table 1). The gut specific expression of 2/20 genes were tested and validated the same (Figure 7).
[0126] Orthologs of the representative TH MIGGS-IRTG (SEQ ID NOs: 76-88) set were identified from transcriptomic resources of an economically important Bt. resistant lepidopteran pest DBM using a combination of reciprocal best Blast analysis (Ward et al. (2014). PLoS ONE
9(7): e101850) and literature curation. Most of these MIGGS-IRTGS were induced in response to the feeding of DBM larvae on induction medium (Wang et al. (2006). J. Biol. Chem. 281(14): 9271-9278), similar to observations with TH larvae.
[0127] It was also demonstrated that most of the MIGGS-IRTGS were induced in another economically important lepidopteran pest, FAW, feeding on plants grown on representative field soil, using a RNA-Seq approach.
[0128] The RNA-Seq approach also identified additional RNAi candidates (SEQ ID NOs: 89-105) belonging to the MIGGS category. Further, orthologs of the expanded representative TH MIGGS-IRTG set (SEQ ID NO: 106-110) were identified from transcriptomic resources of an economically important Coleopteran pest RFB. It was demonstrated that most of the MIGGS IRTGS were induced in response to the feeding of RFB beetles on induction medium (Wang et al. (2006). J. Biol. Chem. 281(14): 9271-9278), similar to observations with the order lepidoptera.
[0129] It was demonstrated that the targeted silencing of 9 out of 20 MIGGS-IRTGS employing bacterially expressed dsRNA protocol (Timmons L. et al. (2001). Gene. 263:103 112.) is insecticidal to TH larvae. Insecticidal activity against TH larvae correlated with the down regulation of target transcripts. It was demonstrated that the targeted silencing of 7 out of 14 MIGGS-IRTGS using bacterially expressed dsRNA protocol (Timmons L. et al. (2001). Gene. 263:103-112.) is insecticidal to DBM larvae. It was demonstrated that targeted silencing of 9 MIGGS-IRTGS are insecticidal to FAW larvae using bacterially expressed dsRNA protocol (Timmons L. et al. (2001). Gene. 263:103-112.). A core set of three MIGGS-IRTGS (SEQ ID NO: 3, 4, and 43) was identified that are efficacious against all three lepidopteran pests TH, DBM and FAW in an orthologous manner and that leaf discs coated with dsRNA against the core MIGGS-IRTGS are insecticidal against TH, DBM and FAW larvae. Further, plastidal expressed dsRNA against the core MIGGS-IRTGS impacted larval growth and survival.
[0130] The following examples are included to demonstrate certain embodiments of the disclosure. It should be appreciated by those of skill in the art that the techniques disclosed in the examples which follow represent techniques discovered by the inventor to function well in the practice of the disclosure. However, those of skill in the art should, in light of the present disclosure, appreciate that many changes can be made in the specific embodiments which are disclosed and still obtain a like or similar result without departing from the spirit and scope of the disclosure.
EXAMPLES
[0131] In certain aspects, work was performed towards the identification, induction, isolation and cloning of the selected M sexta (tobacco hornworm (TH)) MIGGS-IRTGS into a bacterial expression system capable of enabling the cloned genes to produce dsRNA (Timmons L. et al. (2001). Gene. 263:103-112.). Upon ingestion by M. sexta larvae, the bacterially expressed dsRNA is intended to silence, or at least knock-down, reduce, and the like the corresponding MIGGS-IRTG in order to curtail the feeding behavior and/or cause lethal effects in the insect pests. Based upon these results, additional testing was done on other representative insect species to demonstrate and establish for purposes of support wide applicability of the compositions and approaches of the disclosure.
[0132] For MIGGS-IRTG selection, PRR type genes co-regulated by the immune deficiency (IMD) pathway in TH were identified, these genes having been recently summarized (Casanova-Torres and Goodrich-Blair (2013). Insects (4): 320-338; Zhong X, et al. (2012). Insect Biochem. Mol. Biol. 42(7): 514-524); Zhang X, et al. (2015). Insect Biochem. Mol. Biol. 62:38 50; Cao X, et al. (2015). Insect Biochem. Mol. Biol. 62:64-74; Kanost MR, et al. (2016). Insect Biochem. Mol. Biol 76:118-147). Notably, most of the PRR genes selected for this study are abundantly induced or predicted to express in a midgut specific manner (Pauchet Y, et al. (2010). Insect. Mol. Biol. 19: 61-75; Kim and Lee (2014). Front. Cell. Infect. Microbiol. 3: 116; Lee and Hase (2014). Nat. Chem. Biol., 10: 416-424).
[0133] The TH-Transferrin, Arylphorin p subunit, chymotrypsinogen-like protein 1 and few other immunity related genes do not belong to the conventional PRR type immune responsive genes. However, these genes were included as they potentially contribute to the midgut microbial homeostasis through IMD co-regulation (Pauchet Y, et al. (2010). Insect. Mo. Biol. 19: 61-75). Additionally identified were a TH gene indicated to be a valine rich midgut protein critical for the formation of midgut peritrophic matrix and related genes critical for maintaining the structural integrity of the midgut, which are possibly involved, in the gut microbial containment. (Odman-Naresh et al. (2013). PLoS ONE 8:e82015. 10.1371/journal.pone.0082015; Engel and Moran (2013). FEMS Microbiol Rev. 37 699-735). A non-insect gene that encodes for catalase 1 from cassava (Manihotesculanta) was used as a control.
Materials and Methods
[0134] TH larvae required for isolation of IRTG genes and subsequent bioassays were reared as follows: Eggs were procured from Carolina Biological Sciences (Burlington, NC, USA). The eggs were not surface sterilized and used directly for conventional rearing (CR). The larval colony establishment and maintenance was performed employing a Phytatray II (Sigma, MO, USA) unit containing Gypsy Moth diet. (Gunaratna RT and Jiang H (2013). Dev. Comp. Immunol. 39: 388-398).
[0135] Bt.-resistant DBM eggs were procured from Benzon Research (Carlisle, PA, USA) and reared conventionally, as described above.
[0136] FAW eggs were procured from Benzon Research (Carlisle, PA, USA) and reared conventionally, as described above.
[0137] For germ free (GF) rearing procedure, all eggs were surface sterilized with a solution of Tween-80 (polyoxyethylene sorbitan monooleate), bleach, and distilled water as described previously. (Broderick NA, et al. (2009). Environ. Entomol. 29:101-107). The surface sterilized eggs were transferred to Phytatray II (Sigma, MO, USA) unit containing Gypsy Moth diet augmented with antibiotics (500 mg/l each of penicillin, gentamicin, rifampicin, streptomycin). (Broderick NA, et al. (2009). Environ. Entomol. 29:101-107; Gregory R. Richards (2008). JournalofBacteriology. 190, 4870-4879).
[0138] Both CR and GF larvae were reared in an environmental chamber with a 16:8 hours (light:dark) photoperiod at 25 °C, until use. For the induction of a representative set of MIGGS-IRTGS of PRR category, 75 colony forming units (CFU) of DH5a competent cells (Invitrogen, CA, USA) re-suspended in PBS buffer were injected into healthy 2-3 instar M sexta larvae. The larvae were snap frozen in liquid nitrogen and processed for RNA isolation and cDNA synthesis described herein.
[0139] For testing the up-regulation of MIGGS-IRTGS by oral feeding on induction media, first instar larvae were reared on Luria broth agar media plated with a mixture of live E. coli (3 x 107 cells), M luteus (30 pg), and curdlan (30 pg) in 50 pl of H20 (Wang et al. (2006). J. Biol. Chem. 281(14): 9271-9278).
[0140] Total RNA was isolated using RNeasy Mini Kit reagent (QIAGEN, NY, USA) and treated with TURBO DNase (Ambion-Life Technologies, NY, USA) using manufacturer's protocols. One pg of DNase treated RNA was used for cDNA synthesis using iScript cDNA synthesis kit (Bio-Rad, CA, USA). The cDNA was used as a template for amplifying near full length transcripts of the IRTG. Similarly, a control gene from cassava was also amplified. For tissue specific cDNA synthesis the control and treatment larvae were squeezed to isolate hemolymph fraction (HL), dissect midgut (MDG) to obtain rest of the body as described in Pauchet et al. (2010). Insect. Mol. Biol. 19: 61-75). The cDNA template was used for RT-PCR reactions were appropriate using the SuperScript III One-Step RT-PCR system following manufacturers protocol (Thermo Scientific; USA).
[0141] Transcripts of TH, DBM, and FAW MIGGS-IRTGS and non-insect control genes were PCR amplified using PrimeSTAR GXL DNA Polymerase (Clontech Laboratories, CA, USA). The PCR reactions were conducted using the following conditions: denaturation at 98 °C for 30 s, annealing at 55/60 °C for 30 s and elongation at 72 °C for 45 s, for 35 cycles. The PCR products were resolved by agarose gel electrophoresis and stained with ethidium bromide. The transcripts were gel eluted using QlAquick gel extraction kit (QIAGEN, NY, USA).
[0142] Sequence confirmed transcripts were cloned into pCR8/GW vector (Invitrogen, CA, USA) using manufacturer's protocol. The sequence confirmed recombinant pCR8 clones were cloned into L4440gtwy using LR clonase enzyme (Inivtrogen, CA, USA). The L4440gtwy .0 is a modified version of Timmons and Fire feeding Vector and was a kind gift from Guy Caldwell (Addgene plasmid # 11344). (Timmons & Fire (1998). Nature, 395: 854).
[0143] For ingestible RNAi bioassays, sequence confirmed MIGGS-IRTG were cloned into an L4440 feeding vector between two T7 promoters in inverted orientation and transformed into an E. coli bacterial strain carrying IPTG-inducible expression of T7 polymerase, HT115 (DE3). (Timmons & Fire (1998). Nature, 395: 854). Modification of IRTG in this manner was previously demonstrated to induce the expression of dsRNA. (Timmons L, et al. (2001). Gene, 263, 103-112; Kamath RS, et al. (2000). Genome Biol. 2: 1-10.).
[0144] The HT115 (DE3) strain is an RNase III-deficient E. coli strain whose T7 polymerase activity is IPTG-inducible. The HT115 (DE3) genotype is as follows: F-, mcrA, mcrB, IN (rmD-rmE) 1, lambda -, mcl4::Tn1O (DE3 lysogen: lavUV5 promoter -T7 polymerase) (IPTG-inducible T7 polymerase) (RNase III minus), with tetracycline as a selectable marker. (Kamath RS, et al. (2000). Genome Biol. 2:1-10). The standard heat shock protocol for transformation of L4440::IRTG and control construct was used.
[0145] Single colonies of HT115 bacteria containing cloned L4440 plasmids were picked and grown in a 5 mL LB culture with 50 mg/ml ampicillin (Amp). One mL of the liquid culture was saved for plasmid isolation followed by sequence confirmation of the L4440::IRTG clone. The recombinant bacterial clones were grown for 8 hours in liquid culture and were seeded directly on LB plates containing 1 mM IPTG and 50 mg/ml Amp for inducing the dsRNA. (Kamath RS, et al. (2000). Genome Biol. 2: 1-10). Seeded plates were allowed to dry under laminar airflow chamber and incubated at 37 °C temperature overnight.
[0146] For larval bioassay three to five 1-2-instar TH, DBM, and FAW larvae were placed on induced plates containing HT115 (DE3) cells containing the desired L4440::IRTG. Bioassays were conducted testing the TH larvae against MIGGS-IRTGS and controls listed in Table 1.
II+ <
6,
t Al A 0l
-p
I: ,, 9 .* r.
rIr
o r-ctq-o~ M C C tC )" ) N C 00 C> '
o (: dzZz4
0- C: C)z 0
P-iC)
cq IUS
194, C U Uf
[0147] Phenotypic differences in the larval development on L4440::IRTG containing HT115 (DE3) plates were documented and compared with the larval growth on negative and positive controls containing HT115 (DE3) plates. The larval phenotypes for a given treatment with appropriate controls were only considered true and documented if they were reproducibly observed in 2/3 or 4/5 larvae, in at least two independent feeding experiments.
[0148] For sprayable RNAi, a 24-well plate-based bioassay system was developed using a modified cetyl trimethylammonium bromide method of MEGAscript RNAi kit following manufacturer's protocol (Thermo Scientific, USA) for large-scale purification of dsRNA against a given MIGGS-IRTG. Integrity of the dsRNA was determined by electrophoresis on 1% agarose gel and its concentration determined using NanoDrop UV-VIS spectrometer. Leaf discs of 1 cm 2 diameter were detached from Nicotiana benthamiana (for TH) or Arabidopsis Col-WT (for DBM) or wheat cultivar Bobwhite (for FAW) plants grown on field soil were drop inoculated with 0, 4, 8 or 16 pg of purified dsRNA in TE buffer. Air-dried dsRNA coated leaf discs were placed in the bioassay plate containing 1 mL of 1% Murashige and Skoog agar medium per well. Each well contained one leaf disc and was infested with conventionally reared three first instar TH or DBM or FAW larvae. dsRNA coated leaf discs were replaced once every 24 hours and insecticidal activity of each dsRNA measured as a function of larval mortality after five days continuous feeding. The RFB bioassays were conducted using a previously published flour disc assay protocol (Cao et al. (2018). Int. J. Mol. Sci. 19, 1079) with adult beetles.
[0149] For RNA-Seq analysis to test if the MIGGS pathway genes are induced by soil microbiome in FAW, wheat (Triticum aestivum) seeds were surface sterilized and planted in 4.5" pots containing field soil or filled with 4:1 sterile turface:sand mix. Seedlings were grown a growth chamber at for 20 days and infested with ten first-instar FAW larvae per pot. Vigorous larval feeding activity was confirmed and larval samples collected for RNA-Seq analysis. A "pooled RNA-Seq" approach (Rajkumar et al. (2015). BMC Genomics. 16(1): 548) was used to obtain a snap shot of differential FAW gene expression in response to feeding on plants grown on microbe rich (field soil) and microbe depleted (sterile surface) substrate.
[0150] In order to demonstrate additional dsRNA delivery methods a plastidal dsRNA expression system was employed. Since high concentrations of long dsRNAs can be stably produced in plastids (Zhang et al. (2015). Science. 347(6225): 991-994), three of the most potent insecticidal TH MIGGS-IRTGS (SEQ ID NO: 3, 4, and 43) dsRNAs (dsMsPGRP2; dsMspGRP2 and dsMsCHS2) were expressed by plastid transformation in tobacco plants in collaboration with
Plastomics Inc. following a previously published protocol (Zhang et al. (2015). Science. 347(6225): 991-994). Detached leaves of stable transplastomic lines expressing dsRNA against MIGGS targets were fed to TH larvae and assessed for insecticidal activity.
Results
[0151] Bioassays (Figure 1) indicated that the oral feeding activity of TH larvae on L4440::MsPGRP2 and L4440::MsVMP1 containing HT115 (DE3) plates resulted in growth impediment and/or mortality. Contrawise, the larvae growing on L4440::MeCATI containing HT115 (DE3) plates developed normally without any aberrant phenotypes.
[0152] The representative phenotypes of TH larvae at 192 hours post exposure (HPE) to larvae exposed to bacterially (HT115 (DE3)) expressed dsRNA against MIGGS RNAi targets MsPGRP2 (Figure 2A); MsVMIP1 (Figure 2B) and negative control dsRNA against Cassava plant specific gene MeCAT I(Figure 2C). The phenotypic effects leading to larval mortality were usually associated with the development of melanotic reaction, reduced appetite, growth and development (Figure 2A and B).
[0153] Consistent with above, the TH larvae exposed to bacterially expressed negative control MeCATI dsRNA displayed vigorous feeding (area between the arrowheads Figure 3A). In contrast, the TH larvae exposed to bacterially expressed dsRNA against the MIGGS RNAi target MsPGRP2 displayed a curtailed feeding (representative picture, area between arrowheads .0 Figure 3B).
[0154] In general, melanization is a highly conserved immune response and is often associated with microbial infection of insects. (Kim SR, et al. (2005). Insect molecular biology, 14(2): 185-194. doi:10.1111/j.1365-2583.2004.00547). The intensification of melanotic response in TH larvae upon continued exposure to bacterially expressed dsRNA against the MIGGS RNAi targets MsPGRP2 and MsVMP1 containing HT115 (DE3) plates strongly indicates an infection, possibly due to the defective clearance of opportunistic microbes ingested during feeding. Such defective clearance has been previously associated with the perturbation of gut microbial homeostasis. (Packey and Sartor (2009). Curr. Opin. Infect. Dis. 22(3): 292-301).
[0155] Closer observation indicated that the CR larvae feeding on bacterially expressed dsRNA against the MIGGS RNAi targets MsPGRP2 and MsVMP1 displayed discernable mortality starting at day 5, reaching up to 100% and 80% mortality respectively, by day 8 (Figure 4).
[0156] To test if both the incidence and intensity of the observed phenotype could be delayed by clearing gut microbiotas, a GF set of TH larvae were also subjected to the above treatment with appropriate controls, in parallel. Interestingly, that the incidence of larval mortality was not only delayed, but also lower in GF larvae in comparison to CR larvae (Figure 4) was observed.
[0157] The incidence of larval mortality on bacterially expressed dsRNA against MIGGS RNAi targets MsPGRP2 and MsVMP1 plates also correlated with the development of melanotic reaction (Figure 5). Most notably, there was a concomitant delay in the development of melanotic reaction in GF larvae in comparison to CR larvae (Figure 5). All the phenotypes observed were statistically significant at a p-value between 0.001 and 0.05. No significant development of mortality or melanotic reaction was observed in larvae exposed to the bacterially expressed negative control MeCAT IdsRNA (Figures 4 and 5).
[0158] Although MsVMP1 is not directly involved in immune responses, down regulation may abrogate microbial containment, resulting in an infectious phenotype (Figure 2B). Alternatively or in addition, down regulation of MsVMIP1 may have resulted in wounding of the peritrophic matrix (the protective lining of the larval midgut) that also may have contributed to a sepsis mediated infectious phenotype (Figure 2B); this is further substantiated by delayed onset of infectious symptoms in the larvae (CR or GF) exposed to dsRNA against MsVMIP1 in comparison with larvae exposed to dsRNA against MsPGRP2 (Figures 4 and 5). Both defective clearance and defective containment of opportunistic microbes have been previously associated with lethal phenotypes. (Packey and Sartor (2009). Curr. Opin. Infect. Dis. 22(3): 292-301). The schematic representation of dsRNA delivery vectors used for above study is indicated in Figure 6.
[0159] Most of the TH MIGGS RNAi targets disclosed herein (e.g., Table 1) are inducible and have been identified from open access midgut specific immunotranscriptome and/or other datasets (Pauchet Y, et al. (2010) Insect. Mol. Biol. 19:61-75; Odman-Naresh et al. (2013) PLoS ONE 8:e82015; Kanost MR, et al. (2016) Insect Biochem. Mol. Biol 76:118-147; Brummett et al. (2017) Insect Biochem Mol Biol. 81: 1-9; Cao X, et al. (2015) Insect Biochem. Mol. Biol. 62:64-74); Zhong X, et al. (2012) Insect Biochem. Mol. Biol. 42(7): 514-524; Xia Xu et al (2012). Dev Comp Immunol. 38(2): 275-284). A validation study targeting two MIGGS RNAi targets MsHEM and MsSPH3 confirmed their preferential midgut specific manner (Figure
7) when injected with 75 CFU of gram negative E. coi. Further, literature curation confirmed the abundant expression of these MIGGS targets in the insects gut (Table 1).
[0160] Oral feeding of TH larvae on induction media containing a mixture of live E. coli and lyophilized cell wall signatures from gram positive bacteria and fungi (Figure 8) successfully induced (Figure 9) the MIGGS RNAi target genes listed in Table 1. The genes with immunity related function were induced between 24-48 hours post larval exposure to induction media and mostly not detected in the absence of induction (Figure 9A-C). While, the genes essential for midgut structural integrity are expressed under both conditions (Figure 9D). This, clearly suggests that the MIGGS RNAi targets are induced in response to microbes ingested during feeding.
[0161] High-throughput screening of microbe induced MIGGS RNAi targets (Figure 9) using bacterially delivered dsRNA screen identified 3 insecticidal candidates (Figure 10). These MIGGS RNAi targets include Toll receptor (MsToll2), Beta fructofuranosidase 1 (MsSucl) and Beta tubulin (MspTub) genes (Figure 1OD-F). The insecticidal activity was manifested as stunted growth and development, loss of appetite and melanotic reaction (Figure 10D-F) as observed with the positive control MsVATPaseE treatment (Figure 10C) in comparison to the negative control treatment (Figure 10B). The developmental defects due to the bacterially expressed dsRNA exposure resulted in lethality ranging from 60-70% (Figure 11), with MsSucl being the most effective target for killing TH larvae. Mortality rates were statistically significant and comparable to the MsVATPaseE positive control.
[0162] Additionally, a combination of reciprocal best BLAST analysis and literature curation was used to identify the orthologs of TH MIGGS RNAi targets from the DBM transcriptomic resources (Table 2).
tn
o
u r~nCA m nM En En En m En r m r
O0 N
z0 6 -C
w aw -4 co
o U Con p
Oco
1n
-0 U
ot o u "0- C-- =. 9 1)
C)d
~E~u o~~0 C> C~~r zz4
00 0000
o zN.c. z~.o
Cn Qn Cd
>
oo
0 H l I 0 0 I~ QQ
0
[0163] It was demonstrated that 14 MIGGS RNAi target genes identified could also be induced in Bt resistant strain of DBM feeding on the induction media (Figure 12), thus suggesting that oral induction of microbe associated cell wall signatures could induce the MIGGS RNAi target genes in economically important DBM.
[0164] High throughput screening of microbe induced DBM MIGGS RNAi target genes (Figure 12) using bacterially delivered dsRNA screen indicated that 7/15 MIGGS RNAi targets tested showed insecticidal activity against a Bt resistant strain of DBM (Figure 13). The DBM insecticidal targets included PxPGRP2 (SEQ ID NO. 76), PxPGRP2 (SEQ ID NO. 85), PxCHS2 (SEQ ID NO. 87), PxCAC (SEQ ID No. 80), PxIMD1 (SEQ ID No. 77), PxDor (SEQ ID No. 81) and PxpTub (SEQ ID NO. 88). The insecticidal activity was manifested as stunted growth and development, loss of appetite and melanotic reaction (Figure 13D-H) as observed with the positive control MsVATPaseE treatment (Figure 13C) in comparison to the negative control treatment (Figure 13B). The developmental defects due to the bacterially expressed dsRNA exposure resulted in lethality ranging from 53-70% (Figure 14), with PxPGRP2 the most effective target for killing DBM larvae (Figure 14). Mortality rates were statistically significant and comparable to the PxVATPaseE positive control (Figure 14).
[0165] In order to test if our MIGGS RNAi technology could work using multiple dsRNA delivery platforms we tested if dsRNA against the four TH MIGGS insecticidal targets could work in a sprayable format. We used dsRNA against MsPGRP2, MspGRP2, MsCHS2 and MsVMP1 for sprayable RNAi assays (Figure 15) at a concentration of 0, 4, 8 or 16pg of purified dsRNA in TE buffer. Most efficacious leaf disc coated dsRNA was 16 g of dsRNA against MsCHS2 that caused 58% mortality. Similar, but slightly reduced, mortality was observed when insects were fed 8 g of leaf disc coated dsRNA against the same MIGGS targets. Observed mortality rates were statistically significant and comparable to those observed in TH larvae exposed to dsRNA against a known positive control target MsVATPaseE (Figure 16). No statistically significant larvae death was observed when feeding on 4 pg dsRNA (Figure 16), indicating that 8-16 pg dsRNA per leaf disc was required to cause mortality in this assay. In addition to death, TH larvae feeding on dsRNA were characterized by developmental defects, loss of appetite, melanotic reaction and reduced growth compared to negative controls, indicating significant detrimental impact of MIGGs targeting on insect health (Figure 17). The insecticidal TH MIGGS targets MsPGRP2, MspGRP2 and MsCHS2 will be henceforth referred to as core set.
[0166] Similarly, in order to test an additional delivery platform, the core insecticidal MIGGS RNAi targets MsPGRP2; MspGRP2 and MsCHS2 were expressed by plastid transformation in tobacco plants in collaboration with Plastomics Inc. Detached leaves of stable transplastomic lines (Figure 18) expressing dsRNA against MIGGS targets indicated that the TH larvae feeding on leaves expressing dsRNA against MIGGS targets MsPGRP2 (B); MspGRP (C) and MsCHS2 (D) display stunted growth, development, loss of appetite and melanotic reaction in comparison to negative control (A). The insecticidal activity is manifested as significant reduction in mean weights in comparison to negative control (E). The mortality rate was scored on a 0-3 score were 0, 1, 2 and 3 indicated < 0, 25, 50 or > 50 mortality respectively. The transplastomic events confer significant mortality in comparison to negative control (F). Data is average of 6 replicates/treatment (N=24) SEM at p< 0.001(***); p< 0.01(**) and p< 0.05(*). Clearly suggesting that the stably expressed dsRNA against the core MIGGS RNAi targets also impacts larval growth and development.
[0167] Next, we tested if our MIGGS RNAi technology can also work in a sprayable format against Bt resistant strain of DBM. We used the DBM orthologs of insecticidal TH MIGGS core set (PxPGRP2, PxpGRP2 and PxCHS2) and two newly discovered insecticidal DBM MIGGS targets (PxCAC and PxIM4D1) for sprayable RNAi assays at a concentration of 0,1, 0.5 and 0.25 g. We observed lethality ranging from 53-67%, with PxPGRP2 being the most effective target for killing DBM larvae (Figure 19). Mortality rates were statistically significant .0 and comparable to the PxVATPaseE positive control (Figure 19). In a manner similar to that seen for sprayable dsRNA against TH, in addition to death, DBM larvae exposed dsRNA treatments were characterized by developmental defects, loss of appetite, melanotic reaction and arrested growth (Figure 20). Clearly suggesting that dsRNA against the insecticidal MIGGS RNAi targets identified herein when used in sprayable format is also effective against Bt resistant strain of DBM.
[0168] Most of our MIGGS RNAi target gene induction procedures thus far relied upon either direct injection or oral feeding of extraneously supplied microbial signatures. To determine if our MIGGS RNAI target genes are induced under field conditions, we exposed the larvae of economically important lepidopteran pest FAW to wheat seedlings grown on microbe rich and microbe depleted plants (Figure 21). Pooled RNA-Seq analysis indicated that plants growing on field soil caused preferential up-regulation of MIGGS pathway genes in FAW. In total, 100 differential expressed genes were identified, thirty of which were MIGGS pathway related genes (Figure 22).
u
414o 4 4 4
00
C.) - 00 0 0CD 00 4 o ItmN D 0 m efn 000000M C IO~ CIA0CO u0
u C-q 00 - 0~
0' C 0
En 0 0 z
00
0 0 rn o
50c
[0169] Most notably, FAW orthologs of TH insecticidal targets including PGRP2, pGRP2 and IMD were captured in the data set. This discovery indicated that MIGGS pathway genes are up regulated in response to insect feeding on plants exposed to microbes in the field soil.
[0170] Preliminary experiments were performed to screen the insecticidal activity of 17 FAW MIGGS RNAi targets discovered during RNA-Seq (Table 3, above) in which dsRNA against the FAW orthologs of insecticidal TH MIGGS core set (SfPGRP2 (SEQ ID NO. 89), SfPGRP2 (SEQ ID NO. 101) and SfCHS2 (SEQ ID NO. 105) and three newly discovered MIGGS targets SfCTL (SEQ ID NO. 96), SfRC (SEQ ID NO. 103) and SfGAL(SEQ ID NO. 92) from RNA-Seq were fed to FAW larvae at 0, 4, 8 or 16 ig-purified dsRNA in TE buffer. FAW 1st instar larvae were allowed to feed on dsRNA coated leaves following the bioassay described for TH above. Data indicated that FAW larvae (Figure 23) exposed to pure dsRNA against SFCHS2 (B); SFPGRP2 (C); SFPGRP2 (D), SFRC (E), and SFCTL (F) causes reduced growth, development and loss of appetite in comparison to negative control treatment (A) resulting in significant weight reduction (G) and mortality (H) at 8 and 16pg of dsRNA concentration. The rates of mortality was scored on a 0-3 score were 0, 1, 2 and 3 indicated <0, 25, 50 or > 50
mortality respectively. The dsRNA treatments imposed caused statistically significant reduction in mean weights (G) that also translated into significant rates of mortality (H) in comparison to negative control (Figure 23). Screening of insecticidal activity of additional MIGGS RNAi target .0 genes identified from the RNA-Seq dataset indicated that 16 pg of leaf disc coated dsRNA against MIGGS RNAi targets SfITSP (SEQ ID NO. 95), SfAtta (SEQ ID NO. 90), SfCec (SEQ ID NO. 98) and SfHpl0 (SEQ ID NO. 94) caused statistically significant reduction (Figure 24) in mean weights (A) that also translated into significant rates of mortality (B) in comparison to negative control. Data is average of 3 replicates/treatment ±SEM at p< 0.001(***); p< 0.01(**) and p< 0.05(*).
[0171] Experiments with an economically important coleopteran pest RFB were also conducted to test if MIGGS-IRTGS targets identified by a combination of reciprocal best BLAST analysis and literature curation (Table 4) are insecticidal against the order coleoptera.
[0172] Preliminary feeding trails with 1pg of purified indicated that dsRNA against RFB MIGG RNAi targets TcPGRP2 (SEQ ID NO. 107), TcPGRP2 (SEQ ID NO. 108), TcMDGP (SEQ ID NO. 109) and TcCHS2 (SEQ ID NO. 110) is insecticidal to the adult RFB beetles. Significant rates of RFB mortalities were observed (Figure. 25) when scored on a 0-3 scale were
0, 1, 2 and 3 indicated < 0, 25, 50 or > 50 mortality respectively in comparison to the negative control treatment. Data is average of 3 replicates/treatment SEM at ps 0.001(***); p 0.01() and p 0.05(*).
Table 4. List of MIGGS-IRTGS identified from RFB transcriptome resources.
Spodoptera frugiperda Insecti MIGG RNAI Target Genes QueryContigs Putative Insect cidal (SEQ ID NO) _ID Biological Function Midgut Expression Activity
TcPGRPLO DT786101.1 Immunity Yes* No (SEQ ID NOL: 106)
TcPGRP2 XM_965754.3 Immunity Yes* Yes (SEQ ID NO: 107)
TcPGRP2 XM_966587.4 Immunity DNA Yes (SEQ ID NO: 108)
TcMDGP XM_971351 Structural Integrity Yes** Yes (SEQ ID NO: 109)
TcCHS2 EFA10719.1 Structural Integrity Yes*** Yes (SEQ ID NO: 110)
[0173] One possible reason for larval mortality could involve down regulation of MIGGS-IRTGS transcripts upon feeding on exogenously supplied dsRNA against the target genes. Preliminary RT-PCR data indicated that the larval phenotypes (Figure 17) also correlated with down regulation of target transcripts (Figure 26).
[0174] Importantly RNA-Seq analysis indicated that the MIGGS-IRTG pathway targets are induced by soil microbiome indicating that our novel RNAi approach could be effective even under field conditions.
[0175] Given the ease of identification, high specificity, and applicability to diverse pests and delivery platforms, RNAi silencing of the MIGGS-IRTG pathway genes identified, this approach offers an unprecedented potential as a novel pesticidal strategy.
[0176] The translation of these preliminary findings into a pesticidal RNAi technology against economically important pests might lead to sustainable alternatives including but not restricted to the methods described anywhere herein.
[0177] Additionally, the proposed approach will shed more light into understanding the tri-trophic interaction between plants-microbe-insect interactions as it pertains to sustainable insect pest protection.
Sequences
[0178] SEQ ID NOs: 1-14 and 31-44 are representative examples of M sexta-RNAi target gene sequences.
[0179] SEQ ID NO: 15 is non-insect gene sequence that encodes for catalase 1 from cassava (Manihotesculanta).
[0180] SEQ ID NOs: 16-29 are coding region sequences of representative M sexta-RNAi target genes.
[0181] SEQ ID NO: 30 is the coding region sequence of catalase 1 from cassava (Manihot esculanta).
[0182] SEQ ID NOs: 45-70 are 5'UTR and 3'UTR region sequences of representative M sexta-RNAi target genes.
[0183] SEQ ID NOs: 71-75 are the coding region sequences of additional representative M. sexta-RNAi target genes.
[0184] SEQ ID NOs: 76-88 are the coding region sequences of representative P. xylostella-RNAi target genes.
[0185] SEQ ID NOs: 89-105 are the coding region sequences of representative S. frugiperda-RNAi target genes.
[0186] SEQ ID NOs: 106-110 are the coding region sequences of representative Tcastaneum-RNAi target genes.
[0187] SEQ ID NOs: 111-119 are representative examples of Manduca sexta insecticidal dsRNA sequences.
[0188] SEQ ID NOs: 120-126 are representative examples of Plutella xylostella insecticidal dsRNA sequences.
[0189] SEQ ID NOs: 127-135 are representative examples of Spodoptera frugiperda insecticidal dsRNA sequences.
[0190] SEQ ID NOs: 136-139 are representative examples of Tribolium castaneum insecticidal dsRNA sequences.
[0191] > M. sexta-Hemolin (MsHEM); M64346.1 (SEQ ID NO: 1) ATGGTTTCAAAAAGTATCGTCGCTTTGGCTGCGTGCGTCGCAATGTGCGTAGCCCAGCCA
GTGGAGAAGATGCCTGTGCTGAAGGACCAACCCGCTGAAGTCCTCTTCCGGGAGTCTCAG GCCACCGTTCTCGAATGTGTTACCGAGAATGGCGATAAAGATGTCAAATATTCTTGGCAA AAAGACGGCAAAGAATTCAAATGGCAGGAACACAATATCGCCCAGCGCAAAGACGAAGGC AGCCTGGTCTTCCTCAAGCCCGAGGCTAAAGATGAAGGCCAATACAGATGTTTCGCTGAG TCGGCCGCCGGAGTCGCCACCTCCCACATCATCTCCTTTAGAAGGACCTACATGGTCGTA CCTACTACTTTTAAGACTGTAGAAAAGAAACCGGTAGAAGGGTCATGGCTCAAACTTGAG TGCAGCATCCCCGAAGGTTATCCTAAACCTACTATTGTATGGAGAAAGCAGCTTGGTGAA GACGAAAGTATAGCAGATTCTATACTGGCACGTCGTATTACACAATCTCCAGAGGGAGAC CTGTACTTCACGAGCGTCGAGAAAGAAGACGTAAGCGAAAGCTATAAATACGTTTGCGCT GCTAAGTCACCGGCTATTGATGGGGATGTCCCTCTTGTTGGATACACTATTAAAAGCTTA GAAAAGAATACAAATCAGAAAAACGGTGAGCTGGTCCCGATGTACGTCAGTAATGATATG ATAGCTAAGGCCGGAGACGTTACTATGATCTACTGCATGTATGGTGGAGTCCCAATGGCT TACCCCAACTGGTTCAAAGACGGTAAGGACGTGAACGGCAAACCGAGCGACCGCATCACC CGCCACAACAGAACCTCCGGCAAAAGACTGTTCATCAAGGAGACGCTGCTCGAAGATCAG GGCACTTTTACTTGCGACGTGAACAACGAAGTCGGCAAGCCACAGAAACATTCCGTCAAA CTTACCGTAGTCAGTGGACCCAGATTTACGAAGAAACCAGAAAAGCAAGTCATCGCTAAG CAGGGCCAGGACTTTGTAATCCCCTGTGAAGTATCCGCCTTACCGGCCGCCCCTGTCTCC
.0 TGGACGTTCAACGCCAAGCCCATCAGCGGCAGCCGCGTGGTAGCCAGCCCGAGCGGACTG
ACCATCAAGGGCATCCAGAAGTCTGACAAGGGTTATTATGGCTGCCAGGCCCACAACGAG CACGGAGATGCCTACGCTGAGACGCTCGTGATTGTTGCTTAA
[0192] > M. sexta-Serine proteinase homolog 3 (MsSPH-3); AF413067.1 (SEQ ID NO: 2) ATGTTGTTGCTTCTGTATTGTCTTGTGGCGGCCTCCGCGCCGTTCTTTATTGCAGCGGAC
CAAGGCAGCCCTGACCTGCCTTTAGCTACCGAACCACCAACAGAATGCGGAACAATAGCA CCTGATGATAGCTTAGTATTAGATGGGTCCGTTGGTAAAAGTGACAAATTACCTTGGTAT GCTATTATCTACACAACCACCACCCGGCCATACAAGCAGATCGGTGGAGGAACCCTCATC ACTCCTTCAGTAGTAATCTCAGCCGCTCACTGTTTCTGGCGCAATGGTGAGGTTCCATCT AAGGATAATTACGCCGTGGCGCTCGGCAAGACCCATAGTGCTTGGAATAGCCATGCCGAT GTAAACGCTCACAAGTCTGATGTAAAAGAAATACACATACCACCGCAGTTTAAGGGAAGG AACACTAATTATCGGAATGATATAGCAATCGTGGTCATGTCAGACCCTGTGACCTACAAA GTGGACATCCGCCCTATCTGTTTGAACTTCGATGTACAATTTGAAAGACTGCAATTAAAA GACGGCATTATGGGGAAGATCGGCACATGGAATGTAAGTCGTGAGACACTGAAACTATCG AAAACATTAAAAGTGGTGGAGAATCCATACATTGACGCAGCGACTTGTATTAGTGAGTCT CCGGCAAGCTTCAGAAATTCCATCACTGCGGACAAAATATGCATCGGATACGTTAACGGC ACCGGGCTATGTAGAGGTGATGGCGGCGCTGGGGTGGCTTTCCCTAGCCAGGAACAAGGA GTGCAACGTTACTACCTCAGAGGTGTTATATCTACAGCCCATACCAGCGATGATGGCAAC TTATGTGCAGATGGATTTGTAACTGCCACTGCTATAGGCCATCACGAACATTTTATCAAA CAGTTTATAAGCGTTTAG
[0193] > M. sexta-Peptidoglycan recognition protein 2 (MsPGRP2); GQ293365.1 (SEQ ID NO: 3) ATGGCGAGCTTCGCTTTAATAGTTATCCTTAGCGTAATTGGCTTTATATCGGCCTATCCT
AGTCCTGAAGGTTACAGTTCTGCCTTCAACTTTCCATTCGTAACCAAGGAGCAGTGGGGC GGCAGGGAGGCACGCACGTCGACGCCACTCAACCACCCAGTGCAGTTCGTGGTGATCCAC CACAGTTACATTCCCGGCGTGTGCCTCAGCCGGGACGAGTGCGCGCGCAGCATGCGCTCC ATGCAGAACTTCCACATGAACAGTAACGGGTGGAGTGATATTGGATACAACTTCGCTGTC GGCGGTGAAGGGTCGGTGTACGAGGGCCGCGGCTGGGACGCGGTCGGCGCACACGCAGCT GGCTATAACAGTAACAGTATCGGCATCGTGCTCATCGGCGATTTTGTTTCAAACCTCCCG CCGGCGGTGCAAATGCAAACCACACAAGAATTGATCGCAGCGGGCGTGCGACTCGGTTAC
.0 A TCAGGCCCAACTACATGCTCATCGGGCATCGTCAGGTCTCCGCCACTGAGTGCCCAGGA
ACCAGACTCTTCAACGAAATCACCAACTGGAACAACTTCGTGAGGATATGA
[0194] > M. sexta-Beta-1, 3-glucan-recognition protein 2 (MsPGRP2); AY135522.1 (SEQ ID NO: 4) ATGTGGATCAAGAGCGTCTGTTTGTTCGCAACCATTGCGGGCTGCTTGGGCCAGCGAGGG
GGTCCATACAAGGTGCCTGATGCGAAACTCGAAGCTATCTACCCCAAAGGCTTGAGAGTC TCTGTGCCAGATGATGGCTACTCCCTATTTGCCTTCCACGGCAAGCTCAATGAGGAGATG GAAGGTTTAGAGGCTGGCCATTGGTCCAGAGACATCACCAAAGCGAAGCAGGGCAGATGG ATATTCAGAGATAGGAATGCTGAGCTGAAGCTTGGAGACAAAATTTACTTCTGGACTTAC GTTATTAAGGATGGATTGGGATACAGGCAGGACAATGGAGAATGGACTGTTACAGAATTC GTCAATGAGAACGGTACAGTGGTGGACACTAGTACAGCGCCGCCACCAGTAGCACCCGCC GTTTCAGAGGAAGATCAATCGCCAGGTCCTCAGTGGAGACCTTGCGAAAGATCCCTGACT GAGTCCTTGGCCCGCGAACGCGTTTGCAAAGGCAGCCTTGTCTTTAGCGAGGACTTTGAT GGTTCCAGTTTGGCCGACTTGGGCAATTGGACCGCTGAAGTCAGATTCCCTGGCGAACCG GACTACCCGTACAACTTGTACACTACGGACGGCACTGTGGGATTCGAAAGTGGGTCTCTG GTGGTGAGACCCGTCATGACCGAGTCCAAATACCACGAGGGCATCATATACGACCGCCTC GACCTTGAGAGATGTACAGGACAGCTGGGTACGCTGGAATGCAGGCGAGAGAGCAGCGGC GGTCAGATTGTACCACCTGTGATGACAGCTAAACTGGCCACTCGACGCAGCTTCGCGTTC AAGTTCGGCAGGATCGATATAAAGGCGAAGATGCCGCGCGGGGACTGGTTGATACCAGAA CTCAACCTCGAACCTTTAGATAACATATACGGCAACCAGCGATACGCTTCGGGTCTCATG CGGGTCGCGTTCGTGAGAGGAAACGATGTATACGCCAAGAAGCTCTACGGAGGTCCGATA ATGTCCGACGCGGACCCGTTCAGGTCCATGCTGTTGAAGGACAAGCAAGGGTTGGCCAAC TGGAATAATGATTACCACGTCTACTCGCTGCTGTGGAAGCCTAACGGTTTAGAGCTGATG GTGGACGGTGAAGTGTACGGCACCATCGACGCTGGCGATGGCTTCTACCAGATTGCGAAG AACAACCTCGTGAGCCACGCCTCGCAGTGGCTCAAGGGCACCGTCATGGCGCCGTTTGAT GAAAAGTTCTTCATCACTCTGGGTCTTCGCGTGGCGGGTATCCACGACTTCACGGACGGT CCGGGCAAACCTTGGGAGAACAAGGGCACCAAGGCCATGATCAACTTCTGGAACAATCGG TTCCGCTGGTTCCCCACGTGGCACGACACCAGTCTTAAAGTCGACTACGTCAGAGTCTAT GCTCTTTAG
[0195] > M. sexta-Relish family protein 2A (MsREL2A); HM363513.1 (SEQ ID NO: 5) ATGTCCTCTTGTCCAAGCGACTATGATCCCAGTGAATCGTCCAAATCTCCACAAAGTATT
TGGGAGTCAGGAGGATACAGTTCTCCGTCGCAACAAGTTCCTCAATTGACTTCTAACTTA ACAGAATTGTCTGTTGATCACAGCTATAGATACAATGGAAATGGACCATATCTACAGATC ACAGAGCAACCACAGAAATACTTTCGGTTCCGTTATGTTAGCGAGATGGTGGGAACACAT GGATGTTTGCTTGGCAAATCTTATACAACAAACAAAGTTAAAACTCATCCGACAGTTGAA CTCGTGAATTACACCGGTCGAGCCCTGATAAAGTGCCAACTATCGCAAAACAAGAGCGAA GACGAACACCCGCACAAACTGCTCGATGAACAAGACAGAGACATGAGCCACCACGTTCCC GAGCACGGCAGTTATAGAGTGGTATTTGCTGGTATGGGTATAATTCATGCTGCCAAAAAG GAAGTTGCGGGGTGGCTCTATAGAAAATATATACAGCAGAACAAGAATGAAAAGTTTAAT AAGAAAGAGCTCGAAGCGCATTGTGAGAGGATGTCCAAAGAGATCGATTTAAATATAGTT AGACTGAAGTTTAGCGCTCACGATATTGACACTGGCATTGAAATTTGCCGGCCAGTGTTC TCTGAACCCATTTATAATTTGAAGTGTGCGTCTACGAATGATTTGAAAATATGCCGCATA AGCCGTTGTTACGGTAGACCGAGAGGCGGCGAAGATATCTTCATATTTGTCGAAAAGGTC AACAAGAAAAACATCCAAGTTCGGTTCTTTAGACTGGAAAACGGGGAGCGCACCTGGTCA GCGATGGCGAACTTTCTGCTAAGCGATGTTCACCACCAATACGCTATCGCTTTTAGAACG CCACCGTACGTCAATCACCAAATTTCTGAAGACGTGCAAGTTTTTATAGAACTCGTACGC CCTTCAGACGGTAGGACGAGCGCTCCCATGGAGTTCACATACAAGGCTGAGCAAATCTAT AAACAGAACAAGAAACGTAAAACTACTTCGTCGTACTCGTCGCTCGACAGCTCCTCAGGT TCGGCCGGTTCAATTAAAAGCATCAGCGAACTGCCCGCGCCCGTTGTTTTTGCTGAAAAC GTAAGTTTTTTCTATGACACATTACTCATTCTTCAACCCATGACGAATCTATAA
[0196] > M.sexta-Dorsal (MsDor); HM363515.1 (SEQ ID NO: 6) ATGCTTGTGACGTTATGCGGCGGGAACTATAGTGGATTGTCGTTAACAAAAACTAATCAT
TATATGTCACCAAAATCATATGTGCCAGGAAATGGTTATGACGCCGCCGTAATCCTAGGT ACCACGGAGCAGAATGACAGCGAACCCTCAAACTTGAATATTAGTGATGTTTTTGAAGCC ATCACGCTCGCTGATCCGTCGTTCGGCGCGGGCGTGCCGTCGGTAGAGGAGACGATGGCG CACACGCAGCCCCAGCCGCTGCAAATGCCGTACGTGGTCGTCGTGCAGCAGCCCGCCAGC AAAGCGCTCAGATTTCGATATGAGTGCGAGGGCAGATCAGCCGGTTCGATTCCCGGCGCG TCGAGCACGCCCGAGAACCGAACCTTCCCCGCCATCAAGATAATCGGCTACACCGGCACC GTCTCCATCGTAGTGTCGTGTGTCACCAAAGATGAGCCTTGCAGGCCGCACCCACACAAC CTGGTCGGGCGCGACCACTGCGACCGCGGCGTGTTCTCCGTCCGCATCGAGATCACCGAC GAGAATAACGAATACCAGTTTCGGAACCTGGGCATACAGTGCGTCAAGCGGCGCGACATC GGCGAGGCGCTGCGGATCCGAGAGGACCTGCGCGTCGATCCGTTCAAAACCGGCTTCACC
.0 CACCGGAACCACCCGCAAGGCATCGATCTGAATGCAGTGCGGCTCGCGTTCCAAGTGTTC
CTGCCGCACTCCAGCGGCAAGATGCGGCGCACGCTCGCGCCCGTCGTGTCCGACGTCATC TACGACAAGAAGGCCATGAGCGACCTGCTCATCGTGCGCGCGAGCCACTGCGCCGGCCCG GCGCGCGGCGGCACGCAGGTCGTACTGCTCTGTGAAAAGGTGACTCGCGAGGACACCGTG GTGGTGTTCTACCAGGAGGACAACAACCGCGTGCTGTGGGAGGAGATGGCGATCATCATC GTGGTGCACAAACAGTATGCCATAGCGTTCGAGACGCCGCCATACAAGAACCCAAACATT ACTGATAATGTCAATGTACGATTCCAGCTGAGAAGGCTCTCCGACAAGATGACGAGCAAC TCGCTGCCGTTCGAGTACATTCCCGAATACCAAGATTACCCTAGTTACAGGCAGGATAAC TCAGAAAGAAATCCCCAATCGCAGCCAATTACGCACAAGGTAACGGTGGAGGACTTTGAA TCGACAACTAAAAGATATTTTACGCGAAGCACTGACAACAGTAATTACGGTTGGGATGCG GTTCCGGTCACGTACAATGGAAGAAAGAAGGTTTGCTATTGCCCCAAGAGGAGCTAA
[0197] > M. sexta-Spatzle (MsSPZ1A); GQ249944.1 (SEQ ID NO: 7) ATGGCCTGGATCCAGCATTTACTCGTTTGGCTCTTCGTTATGTCAACATCAGCATACAAA
TGCAAAGACTGCTTCAGTTTCGCATCACAATATCCGTCGTACGATAGTCAAGTATACGAA CAACCTGACAGACGGATAGCGGGACGGTCAGCACAATACGAACATTTAAGAACAAACGAG AGGTCTCTCCCGGTCTACAGCGAGACCCAGAGGATACAAGCAGAAGAGAGAAGAAGACAC AGTTCGAGACTAGAAGAACCGAGACAACGTGCTGAGAATGGTTCATATAAGATATTGAAT AACCCTCCGAAACCCTGTATTACTAATAGGAGAAGTCAAATTGATTCGTCGAATGATAGG GTAGTGTTCCCCGGTCCGACTTCAGAAAGGTCGTACGTACCCGAAGTGCCAGAGGAATGC AAGAAAATCGGCATATGCGACAGTATACCGAATTACCCAGAAGAACACGTAGCTAATATT ATATCTCGACTTGGAGACAAAGGAAAAGTATTACAAATAGACGAACTGGACGTATCAGAC ACTCCAGATATCGCCCAGAGGTTGGGTCCGCAGGAGGACAACATGGAACTATGTAGCTTT AGAGAAAAGATTTTTTACCCCAAGGCAGCGCCAGACAAAGATGGAAATTGGTTCTTCGTT GTGAATTCAAAAGAAAACCCAGTACAGGGTTATAAAGTTGAAATTTGCGACCGTCAGCAA TTACCATGCGCGGAGTTCGCGAGCTTCCAACAGGGATATGAAGCGAGGTGCATCCAGAAA TACGTTCGCCGGACCATGTTGGCGTTGGATCCCAAGGGTCAGATGACCGACATGCCCCTT AAAGTGCCCAGCTGTTGCTCATGCGTGGCCAAATTGACAATCATATGA
[0198] > M. sexta-Toll receptor (MsTOLL); EF442782.1 (SEQ ID NO: 8) ATGCAGGCTCGGCGGTGGTGCGCGGCACTGCTATTAATGCAGATGCTGAGCTGGCTCGGA
.0 G TCAGTGGACACTTACCGCGTCCCGAGTGCGCGCCAGCCGCAGATTGCCAACTTATACGA
GACAACATAATCGATGGATATGCACAATTCTACTTCAACGTATCAGGACATGAAGTGAAA TTTGAACATTACATCGGAAACGACTTCGATGTCGAATTGTCATGCAATTACATCGCCATG GACAACGCAATGCTGCCGCGGTTCTCAACGACCTTTTCAGTCAACGTAATAGTGGTTAAA GAATGTGCTTTGCCAAGAAGTGGGTCAATCGATGCCGCTGTCGCTGCACTTAATATCAAC GTTTTGACGGAGCTGACTCTGGACAAATTCCTAGAGCCGGCGGTGATCACGCGCGCACAT CTTACCAGTTTACAACGACTAGAGAGGCTGGAGCTACACGGTAACTCAAACACAAGCCTC GCCCCCGGCGCACTGGCCGCGCTCTCCGCCGCGTCCGCACTGAAATGTCTTGTATTGCAT GCAGTACGCGTGCCCGCCGCTGACCTGGCGCGCTTGCCGTCGTCACTGCAAGAACTAGCG TTGTTGGATGTGGGCGCTGCGAGTATGCATTTAGATTCATCGGTTAATTTGACGTCACTC TTCGTAATCGATACACATTATCCTGTCGTCGTGAATGTGAGCAACGCCGTTGCGCTCAGA GACTTGCACATAAATACCCCAAGTACTGTGTTGACCGAAGACGTGCTCCCGTCGTCACTC AACTCACTTGAACTAGAGGGGTGGAACGAAACGCATCCGGTGCCTAAGACACGTTGTGTA CTACTTAAGGAACTTAATGTAATCGGCACCGACAATGATGCCTATCCGGTGACTCTCCCG GACGAGTGGCTGTCCAACTGCGGACAGCTGAGGGATCTTGAAATGATCTCCGTGCCGATT AGCGCCGTACTTCCGGCGCGGATGCTGGCTAACGCAATTAGGCTTGAAACGATTACTATC TGGAACTGTAACCTGACCGCGTTGCCGTCGGGCCTGTTAGACGACACGCTGAACCTCGCC ACACTCGACTTGTCCAACAATCAACTTGCATCGTTGCCCAGAAAATTATTTGAGCACACG AAGTTACTACGCAGTCTCATACTATCGAACAATCAGCTGACGAGCGAGGTAGTGTGGACG CTGTCGACCGTCACCTCGCTTGTTGAACTAAAGCTCAGCAATAATAACCGCATAGGCGAC TTATGTTCCCACGACTCAGTGTCAGCAGGTCCCTCACCACTGAGTTCACTGACGGGGCTA AACTATCTCCATCTGAGCAACACAGGAGTATCACACGTGTGCTCCGACTGGCGAGAAAAG CTAACCTACCTCACGAATCTCAACTTAAGGGACAACCCCATCACTCTCTACAATTTGGCG GATCTACAGTTTCGTCGAATCTGGGTGGACGCTCGTGTGTATTTAGGACATTTCAAGCAG CAGTTTACGCGCACAGATTACGAACTCGCCAGTAATAATAACACTGAGGCGGTCGTAACT TTATCCGGTTCGTTAGAATGCGACTGCAATTCATACTGGGCAGCACAGGTGTTCCGTATG AAAGCTTGGCAGGCATCCAGCTCCATGATATATTGTGAAAAAAAACCGGTCATTGAGGTG GATCCCGACACCTTTACCTGCCTTGAGCCAGCGAAGTGTGCTGCGCTGGCGGATAGTTGT ACGTGCCGTATCCGCGACGACATTCAGTACAAACAAGTGGTCGTTGTGCACTGCACCGGA CTCGCCGAGTTCCCGCGCCTGCCACTAACCACGGACAAGTGGATCCTGCACCTGCCCCAC AACAACATATCATACCTCGCCGCGGCCGACGTATCGCCGAACATCGTGGAACTCGATCTA AGAAACAATTCAATCAAGAATATCGACGTACAGGCATCAGCAAAACTAGCCTTCGTCCGG CTGCAATTGGGTGGTAACCCGATCGAGTGTGACTGCGAGGCGTTGAAGCTGCTGGCGCCG
.0 C TGCTCAAACCTGACTCAAAGCTGCTTGACAGAAAGGACGTGAAATGTGAGAACGACGCG
CAGATTACCTTGGCGATGCTGAAATTATGTACTAAGTCATCCAATGGGCTGATGTACTTG CTGTTCCTGTTGCTGTTACTCGCATTTGTCGTAACCGGACTGCTCGCACGAACCGCAATT CGCCTGCGCATCAAAATGATCCTCATGAGACTGGGTTGGATGTCGAGACTACTGGAGCCC GCGGACGACGATCGCCCGTACGACGCGTTTGTGTCTTTCGCACACGAGGATGAGGAGCTG GTGATGGAGCAGCTGGCGGCACGGCTCGAGAGCGGCTCGCGGCCGTACCGACTGTGTCTG CACTACCGCGATTGGGCGCCAGGCGAGTGGATCCCGGCGCAAATAGCGGCTTCGGTGCGG GCCTCTCGGCGCACGGTGGCGGTTGTGTCGGCGCACTACTTACAGTCGGGCTGGGCGCTT GCCGAGATCCGGGAGGCGACCGCAGCCTCGCTGCAGGAAGGCATGCCACGTCTCATCATC GTGCTGCTCGACGAGACCGACCGGTTGATGCTCGATATAGACCCTGAGTTGCACGCCTAT GTGCGCAACAATACCTACGTGCGCTGGCATGATCCATGGTTTTGGGAGAAGCTGAAGCAG GCGCTGCCTCCACCGCGGGAACAACGGTCGCCAATAGCTCCGTCGCTACCAGCGCTGGCG CTGTCCCATGACAGCCTAACTCTGCGCACGTACTCTCCCAGAGAAAGTGACCCAGCGCCC GCTGCTAAGCCGGCGCACACTCCGCACGAGACGGACGAGCCAGCGCCGGGCGCGAGTCCT TGCTACAAATGA
[0199] > M. sexta-Scolexin A (MsSCA1); AF087004.1 (SEQ ID NO: 9) CGGCAGTCGGTTGTGTTGGCAGTGGCGGCGGTGCTCTTCGGGTGCGCGTGCGCAGCGCCC
AATCCTGGCGCCAACGACATACAACTTAATCAAAAATTAAGTATCGAAGCTAAGGGGGCA AAGCAGCCAATTGATACGAGGGCAGTGAAGGAACGGTATCCATACGCAGTTCGGAGTTTC GGAGGCTTCTGCGGAGGAACCATTATCAGTCCCACCTGGATCCTGACCGCCGGCCACTGC TCGATACTCTATGCGGGGAGCGGCCTACCGGCCGGCACCAACATTACCGAGGTATCTAGC TTGTACCGCTTCCCCAAGCGGCTCGTCATACACCCGCTCTTCTCCATAGGACCCGTCTGG CTCAACGCTACGGAGTTCAACCTCAAACAGGCGGCTGCACGATGGGACTTCTTGTTGATA GAACTGGAGGAACCGCTGCCGTTGGACGGCAAGATCCTGGCGGCTGCGAAGCTCGACGAC CAGCCCGACCTCCCCGCAGGCCTCGACGTGGGCTATCCGAGCTACAGCACCGACACCTAC GAGGCTAAGATACAAAGCGAGATGCACGGAAAGAAGCTTTCGGTTCAATCTAACGAGGTG TGCTCGAAGCTAGAGCAGTTCAAGGCGGAGGACATGTTGTGCGCCAAGGGACGTCCACCG CGATACGACTTCGTCTGCTTCAGCGACAGTGGCAGTGGGCTAGTAGACAACAATGGTCGC CTAGTCGGCGTGGTGTCGTGGGCCGAGAACAACGCTTTCGAGTGCCGCAACGGCAACCTG GCGGTCTTCTCGCGAGTGTCCAGCGTACGCGAGTGGATCCGACAAGTCACCAACATATAA
[0200] > M. sexta-Hemolymph proteinase 18 (MsHP18); AY672794.1 (SEQ ID NO: 10) ATGGTTTATATTTTAATAATTTTAGTGATTTGCAATTTTAGTTGTATTAGTTGTCAGTCC
GGGACAGTGGAAAGCAGGATTCATTTTAAAGATGAAGGGCCGGAATGTTATGATGCAAAT AAAAAGGGCACCTGTGTTAGTGCTCACAGATGCCTTGATGTAGTTAGAAAACTTAAAGAC GGAGAGAAACCCACGATATGTGGCTACCAAGGCACGGAACCAATGGTGTGTTGCACAGAC TGTACTCTGGTTGATAATATTAGTAATTTGGTCGTAAGTTCCATATCCGGGTACCTGTGG AAGGATGGTCAGAAAGCGTGGGACAAATGTCTGGAATACGTTGACAAGCTGTCGTACCCA TGCGCTTCAACCTACTCCCACTACCTCAGCTCCGTTTGGGAGAAAGATAAGGAGTGCAGT ATGGTTCAGTTTGTTGGCGTGAGGCGATTCGCCTCGTATAACGGACAACCGGCGAAACGG AACGAGTACCCTCACATGGCTCTGCTCGGCTACGGCGACGACCAGGAGACGGCGCAGTGG CTCTGCGGCGGCTCAGTGATCAGTGATCAATTCATCCTCACGGCTGCACACTGCATCTTT ACAAATCTATTGGGTCCAGTACGTTTCGCAGCGCTAGGAATACTGCAGCGATCGGATCCA GTAGAGTTATGGCAAGTTTACAAGATCGGCGGCATAGTTCCCCATCCGCAGTATAAGTCA CCTATCAAGTACCACGACATTGCTCTCCTGAAGACTGAAAACAAAATAAAGTTTAATGAG AACGTGCTGCCAGCGTGTTTGTTCATAGAGGGCAGAGTGGGTGGGAGTGAGCAGGCTAAA GCGACCGGTTGGGGCGCGCTTGGACATAAACAGACGGCAGCTGACGTACTGCAAGTGGTT GACCTTCAAAAGTTCAGTGACGAAGAGTGCGGAAGTACCTACCGTCCTTACCGGCATTTG CCTCAAGGCTACGACAGCGCCACGCAGATGTGCTACGGCGACAAGGGAAAACTGAATATG GACACCTGTGAGGGCGACAGCGGCGGTCCTCTACAGTTCCAAAACTCCTCGCTCCTCTGC ATACACATAGTAGCGGGAGTGACGTCATTCGGCGACGCGTGCGGGTTTGCGGGCGGCGCC GGGATGTACACACGAGTGTCGTACTATATTCCCTGGATCGAGAGCGTTGTATGGCCGTGA
[0201] > M. sexta-Transferrin (MsTRN); M62802.1 (SEQ ID NO: 11) ATGGCTTTGAAACTTTTAACTTTGATAGCCCTGACTTGTGCGGCTGCGAATGCAGCTAAA
TCTTCATACAAACTATGCGTGCCAGCAGCATACATGAAGGACTGCGAGCAGATGCTTGAA GTACCCACGAAGTCTAAAGTGGCCTTGGAATGTGTACCGGCTAGAGACAGGGTGGAATGC CTCAGCTTTGTTCAGCAGCGACAGGCGGACTTCGTCCCCGTCGACCCTGAGGACATGTAC GTGGCCTCCAAGATCCCCAACCAGGACTTCGTCGTCTTCCAGGAGTACAGGACTGATGAA GAGCCTGATGCGCCATTCCGTTATGAAGCCGTTATTGTGGTTCACAAAGACCTACCCATC AACAACTTGGATCAGCTGAAGGGACTGAGGTCTTGCCACACCGGAGTCAATCGTAACGTC GGGTACAAGATCCCACTAACGATGTTGATGAAACGTGCCGTGTTCCCGAAAATGAACGAC CACAGCATTTCGCCGAAAGAGAACGAACTGAAAGCGCTATCGACGTTCTTCGCAAAGTCG
.0 TGCATCGTCGGCAAATGGTCGCCTGACCCCAAAACCAACTCGGCTTGGAAATCACAATAC
AGCCATTTGTGTTCAATGTGCGAACACCCGGAGCGTTGTGACTATCCCGACAATTACAGC GGGTACGAGGGCGCGTTGAGATGCCTCGCCCACAACAACGGGGAGGTCGCGTTCACCAAA GTCATATTCACACGTAAATTCTTTGGGCTTCCAGTAGGTACCACTCCAGCGAGTCCATCA AACGAAAATCCCGAAGAGTTCAGATATCTCTGCGTGGACGGATCTAAAGCCCCCATCACT GGCAAGGCTTGTTCATGGGCTGCCAGACCTTGGCAAGGACTGATCGGTCACAATGACGTA CTTGCCAAACTCGCTCCGCTCAGAGAGAAGGTTAAGCAACTTGCTGATTCTGGTGCAGCT GACAAACCGGAGTGGTTCACCAAAGTCCTTGGTCTATCAGAGAAGATCCACCATGTCGCT GACAATATCCCAATCAAGCCCATCGACTACCTGAACAAGGCTAACTACACGGAGGTCATT GAAAGAGGACATGGAGCTCCCGAGCTGGTCGTCAGGCTATGTGTGACGTCAAACGTGGCA TTATCTAAGTGCCGGGCTATGTCCGTGTTCGCATTCAGTAGAGACATCAGGCCGATCCTA GACTGTGTTCAAGAAAACAGCGAAGATGCCTGTCTTAAGAGCGTCCAAGACAACGGTTCA GATCTTGCCTCAGTAGACGATATGAGAGTAGCTGCAGCGGCTAAGAAGTACAACTTACAT CCAGTTTTCCACGAAGTGTATGGAGAGCTAAAGACGCCCAACTACGCAGTGGCTGTTGTC AAGAAGGGCACTGCCTACAACAAGATCGACGACTTAAGGGGAAAGAAATCTTGCCACAGC TCTTACAGTACTTTCAGCGGTCTGCACGCGCCTCTCTTCTACCTTATTAACAAGAGGGCC ATTCAATCTGACCACTGCGTGAAGAACTTGGGAGAATTCTTCTCAGGCGGATCTTGCTTG CCTGGTGTCGACAAACCCGAAAACAACCCAAGCGGTGATGATGTGTCTAAATTGAAGAAG CAATGTGGATCCGACAGCAGCGCTTGGAAGTGCTTGGAAGAGGACAGAGGAGACGTCGCA TTTGTTTCAAGTGCCGATCTGTCCCACTTCGACGCCAACCAATACGAGCTGCTCTGCCTG AACCGCGACGCTGGCGGTAGAGATGTTCTCTCCAGTTTCGCCACTTGCAACGTCGCCATG GCCCCGTCCAGGACCTGGGTGGCTGCGAAGGACTTCCTGTCTGACGTATCTATCGCCCAC ACACCATTGAGCCTCGCCCAAATGCTCGCTACGAGACCTGACCTCTTCAACATTTACGGA GAGTTCTTGAAGAACAACAATGTTATTTTCAATAATGCCGCTAAAGGCTTAGCAACAACT GAGAAACTTGACTTCGAGAAGTTCAAGACCATCCACGACGTCATCTCTTCATGTGGTCTC GCCTAA
[0202] > M. sexta-Arylphorin P subunit (MsARP); M28397.1 (SEQ ID NO: 12) ATGAAGACTGTCATAATCCTAGCGGGGTTGGTGGCCCTGGCCCTCGGCAGCGAAGTGCCT
GTCAAGCACTCCTTCAAAGTTAAGGATGTTGATGCGGCTTTCGTCGAACGTCAAAAGAAG GTCTTAGATCTTTTCCAAGATGTCGACCAAGTAAATCCTAACGATGAGTACTACAAGATT GGCAAGGAATACAACATCGAGGCTAACATCGACAATTACTCGAACAAGAAGGCCGTCGAA GAATTCTTGCAGTTATACAGGACAGGTTTCTTGCCTAAGTACTATGAATTTTCACCCTTC
.0 TATGACAGACTAAGGGACGAGGCCATTGGTGTTTTCCACCTCTTTTACTACGCTAAAGAT
TTTGATACGTTCTACAAATCTGCCGCATGGGCGCGTGTGTACCTCAACGAAGGACAGTTC TTATACGCCTACTACATTGCTGTGATTCAGCGTAAAGATACTCAGGGCTTCGTTGTACCA GCACCGTATGAAGTCTACCCTCAATTCTTCGCAAACTTGAACACTATGCTCAAAGTCTAC CGTACCAAAATGCAGGATGGAGTTGTTAGTGCCGATTTAGCTGCACAACACGGCATCGTA AAGGAGAAAAACTACTACGTATACTATGCCAATTACTCCAACTCATTAGTGTACAACAAC GAGGAACAGAGACTGTCGTACTTCACTGAGGACATCGGCTTGAATTCGTACTACTACTAC TTCCACTCTCACTTGCCTTTCTGGTGGAATTCTGAGAGATACGGAGCACTAAAATCGCGC CGTGGTGAAATCTACTATTACTTCTATCAGCAATTAATTGCACGTTATTACTTTGAACGT CTCTCGAACGGCCTGGGTGACATTCCCGAATTCTCATGGTACTCACCAGTCAAGTCTGGC TACTATCCACTGATGTCTTCTTATTACTACCCCTTCGCTCAAAGGCCCAACTACTGGAAC GTGCACAGCGAAGAAAACTACGAGAAAGTACGATTCTTGGACACGTATGAAATGTCATTC CTTCAGTTCCTCCAAAACGGACACTTCAAAGCGTTTGACCAGAAGATTGACTTCCACGAT TTCAAAGCTATCAACTTTGTTGGAAACTACTGGCAAGATAATGCTGACCTGTACGGTGAG GAAGTTACTAAGGACTACCAACGTTCATATGAAATTATAGCCCGCCAAGTGCTTGGTGCT GCACCTAAACCATTCGACAAGTACACATTCATGCCCAGCGCTTTAGACTTCTACCAGACG TCTCTGCGTGACCCAATGTTCTACCAACTTTACAACAGAATTCTGAAGTACATATATGAG TACAAGCAGTACCTGCAACCGTACTCTTCAGAAAAACTGGCATTCAAGGGTGTCAAGGTG GTCGATGTTGTAGTAGACAAACTGGTTACCTTCTTCGAGTACTACGACTTTGATGCGTCC AACAGCGTTTTCTGGAGCAAAGAGGAGGTTAAATCTAGCTACCCCCATGATTTCAAGATC CGTCAGCCACGCCTTAACCACAAGCCATTCTCTGTCTCTATCGACATCAAATCTGAAGCT GCCGTTGATGCCGTTGTCAAGATATTCATGGCACCTAAATACGACGATAATGGATTCCCT CTGAAATTAGAAAACAACTGGAACAAATTCTTCGAGCTGGACTGGTTCACATACAAATTT GTTGCTGGTGACAACAAAATCGTGAGGAACTCAAACGACTTCTTGATCTTCAAGGACGAC TCTGTTCCCATGACTGAGTTGTACAAATTATTAGAACAAAATAAGGTTCCACACGACATG TCCGAGGATTACGGCTACCTGCCTAAAAGACTGATGCTGCCAAGAGGTACTGAGGGTGGT TTCCCATTCCAGTTCTTCGTTTTCGTATATCCATTCAACGCTGACAGCAAAGATCTTGCA CCGTTCGAGGCCTTCATCCAGGACAACAAACCTTTGGGCTATCCATTCGACCGTCCCGTT GTTGACGCTTACTTCAAGCAACACAACATGTTCTTCAAGGACGTCTTCGTATACCATGAC GGCGAGTACTTCCCGTACAAGTTCAATGTTCCTTCCCATGTGATGCACTCAAACGTTGTT CCTAAACACTGA
[0203] > M. sexta-Chymotrypsinogen-like protein 1 (MsCTL1); AM419170.1 (SEQ ID NO: 13) ATGTACGTGAAAGTAGCACTTCTGTTGGTAGCCCTCATTGCTGGGAGCTGGGCCTTCCCA
AAGCTCGAAGATGAGCAGGACATGTCCATCTTCTTCACGCAGCTCGATTCGAGCGCGCGT ATCGTGGGTGGTACCCAGGCCCCCAGCGGAAGTCACCCTCACATGGTGGCGATGACCACC GGTACCTTCATCAGGAGCTTCAGCTGTGGAGGCTCAGTTGTCGGTAGACGTTCCGTTCTG ACTGCGGCTCATTGCATCGCTGCTGTTTTCAGTTTCGGTTCCCTCGCCAGTACCCTCCGC TTGACGGTCGGCACCAACTTCTGGAACCAGGGAGGCACCATGTACACCGTCGCTCGCAAC ATAACCCACCCCCACTACGTCTCTGCGACCATCAAGAACGACATCGGTCTGTTCATCACT CACAACAACATCATCGACACGACTGTCGTCCGCAGCATCCCTCTTAACTTTGACTATGTG CCCGGTGGTGTTCTCACTAGAGTCGCCGGATGGGGCAGGATCAGGACCGGCGGTGCCATC TCTCCCTCTCTGCTGGAGATCATTGTGCCTACTATCAGTGGAAGCGCATGCGTAGCCAGT GCAATCCAAGCTGGCATCGATCTGAACATGAGACCACCTCCCGTCGAGCCTCACATCGAG CTGTGCACCTTCCACGGTCCTAACGTAGGCACTTGTAATGGTGACTCCGGCAGCGCTCTT GCCCGCCTAGACAACGGCCAGCAGATCGGTGTGGTATCGTGGGGCTTCCCGTGCGCACGC GGCGGTCCCGACATGTTCGTCAGGGTCAGCGCCTACCAATCCTGGCTGCAGCAGAGCATC GTATAA
[0204] > M. sexta-Valine Rich Midgut Protein (MsVMP1); NCBI accession number not assigned as yet (SEQ ID NO: 14) ATCATTGACGGACCTTCCGTTGGACCNGCCATCATCGGCGCTGGAGACATCGCTGTCGGC
CCTGCTATCGTCGACTTCCCTTTCCCCGACGGCGGTGCCGTGTCTGCCCCCGTTGAGCCT TCCCCCATCGCCATCGGACCCGCTATCGTCGGTGAATCCCCTATCTCCGTCGGACCTGCC ATCGTTGAGGCCGGAGACATCGCTGTTGGACCCGCTATCATCGACTTCCCCCTTCCCGAC GGTGGCGCCGTGTCCGCCCCCGTTGAGGTTTCTCCCGTCGACTCCGTCGTCGTCGGCCCT GCCGCCGGCTCTCAGAGCTCTCCCCTCGTCCAGATCATCATCAACGTTAAGGCCCCCGCT GGTGCCGGCCCCGTTGTCGATGCCGTCGCTGACAAGCCCATGGACATCATTGATGTTATG CCCGTCGTCGACCCTGCTGATTTCGTGGACCTCACCCCCGTTGTAGAGCCTGTAGAAGTC GTCGACATTGTCGATGTCATGCCCGTGGTTGACCCCATCAACATCATCGATGTTATGCCT GTTGTTAAGCCCGTAAACCCCCTTGCCCGTTCTTAAGGG
[0205] > M. esculanta-Catalase 1 (MeCAT1); AF170272 (SEQ ID NO: 15) ATGGATCCTTGCAAGTTCCGTCCATCAAGCTCAAACAATACCCCCTTCTGGACCACCGAT
GCTGGTGCTCCAGTATGGAACAACAATTCCTCCATGACTGTTGGAACCAGAGGTCCAATC CTTTTTGGAGGACTATCATATGATAGAGAAACTTGCCAACTTTACCAGAGAGAGGATTCCA GAGCGTGTCGTCCATGCTAGGGGAATGAGTGCAAAGGGCTTCTTTGAAGTCACCCACGAT GTCTCTCACCTTACTTGTGCTGATTTCCTTCGAGCCCCTGGAGTTCAAACCCCTGTCATC GTCCGTTTCTCCACTGTTATCCACGAGCGTGGCAGCCCTGAAACACTCAGGGATCCTCGA GGTTTTGCGACTAAGTTCTACACCAGAGAGGGCAACTTTGATATTGTGGGAAACAACTTC CCTGTCTTCTTCATCCGTGATGGAATAAAATTCCCAGATGTGATACACGCTTTTAAGCCC AATCCCAAGTCTCACATCCAAGAATACTGGAGGATCTTTGACTTCTTATCACACCATCCT GAGAGCTTGAGCACCTTCGCCTGGTTCTTCGATGATGTTGGAATTCCCCAAGATTACAGA CACATGGAAGGTTTCGGTGTTCACACCTTTACTTTCATCAACAAGGCTGGAAAAGTAACC TACGTGAAATTTCACTGGAAACCCACTTGCGGGGTCAAGTGTTTGATGGATGATGAGGCA CTTAAGATCGGAGGTGCCAACCACAGCCATGCTACGCAGGATTTATACGACTCCATTGCC GCTGGCAACTATCCTGAGTGGAGACTCTTCATCCAGACAATGGATCCAGCTGATGAAGAC AAATTCGACTTTGATCCACTTGATATGACCAAGATCTGGCCTGAGGATATTTTTCCTCTA CAGCAAATTGGCCGTTTGGTCTTGAACAGGAACATCGATAACTGGTTTGCTGAGAATGAA ATGCTCGCATTCGACCCTGGTCATATTGTTCCTGGCATTCACTATTCAAACGACAAGTTG TTTCAGCTCAGAACCTTTGCATATGCTGACACTCAGAGGCACCGTCTCGGACCCAACTAT AAGATGCTCCCTGTTAATGCTCCCAAGTGTGCTTATCACAACAATCATTACGATGGTTTC ATGAATTTCATGCACAGGGATGAGGAGGTGGATTACTTCCCATCCAGGTATGATCCAGTT CGCCATGCTGAGAGAAGCCCCATTCCTAACGCTATCTGTAGTGGAAGGCGTGAAAAGTGC GTCATTGAAAAGGAGAACAATTTCAAGCAACCTGGAGAGAGATATCGATCCTGGGCACCT GATAGACAAGAAAGATTCCTGTGCAGATTGGTTAACGCCTTATCAGAGCCACGTATCACC TTTGAGATTCGCAGTATCTGGGTCTCTTACTGGTCTAAGTGCGACGCGTCTCTGGGTCAA AAGCTGGCTTCTCGTCTCAACGTGAGGCCAAATATATGA
[0206] > M. sexta-Hemolin (MsHEM); M64346.1 (SEQ ID NO: 16) GGCAAAGAATTCAAATGGCAGGAACACAATATCGCCCAGCGCAAAGACGAAGGCAGCCTG
GTCTTCCTCAAGCCCGAGGCTAAAGATGAAGGCCAATACAGATGTTTCGCTGAGTCGGCC GCCGGAGTCGCCACCTCCCACATCATCTCCTTTAGAAGGACCTACATGGTCGTACCTACT ACTTTTAAGACTGTAGAAAAGAAACCGGTAGAAGGGTCATGGCTCAAACTTGAGTGCAGC ATCCCCGAAGGTTATCCTAAACCTACTATTGTATGGAGAAAGCAGCTTGGTGAAGACGAA AGTATAGCAGATTCTATACTGGCACGTCGTATTACACAATCTCCAGAGGGAGACCTGTAC TTCACGAGCGTCGAGAAAGAAGACGTAAGCGAAAGCTATAAATACGTTTGCGCTGCTAAG TCACCGGCTATTGATGGGGATGTCCCTCTTGTTGGATACACTATTAAAAGCTTAGAAAAG
.0 AATACAAATCAGAAAAACGGTGAGCTGGTCCCGATGTACGTCAGTAATGATATGATAGCT
AAGGCCGGAGACGTTACTATGATCTACTGCATGTATGGTGGAGTCCCAATGGCTTACCCC AACTGGTTCAA
[0207] > M. sexta- Serine Proteinase homolog 3 (MsSPH-3); AF413067.1 (SEQ ID NO: 17) ATGTTGTTGCTTCTGTATTGTCTTGTGGCGGCCTCCGCGCCGTTCTTTATTGCAGCGGAC
CAAGGCAGCCCTGACCTGCCTTTAGCTACCGAACCACCAACAGAATGCGGAACAATAGCA CCTGATGATAGCTTAGTATTAGATGGGTCCGTTGGTAAAAGTGACAAATTACCTTGGTAT GCTATTATCTACACAACCACCACCCGGCCATACAAGCAGATCGGTGGAGGAACCCTCATC ACTCCTTCAGTAGTAATCTCAGCCGCTCACTGTTTCTGGCGCAATGGTGAGGTTCCATCT AAGGATAATTACGCCGTGGCGCTCGGCAAGACCCATAGTGCTTGGAATAGCCATGCCGAT GTAAACGCTCACAAGTCTGATGTAAAAGAAATACACATACCACCGCAGTTTAAGGGAAGG AACACTAATTATCGGAATGATATAGCAATCGTGGTCATGTCAGACCCTGTGACCTACAAA GTGGACATCCGCCCTATCTGTTTGAACTTCGATGTACAATTTGAAAGACTGCAATTAAAA GACGGCATTATGGGGAAGATCGGCACATGGAATGTAAGTCGTGAGACACTGAAACTATCG AAAACATTAAAAGTGGTGGAGAATCCATACATTGACGCAGCGACT
[0208] > M. sexta- Peptidoglycan recognition protein 2 (MsPGRP2); GQ293365.1 (SEQ ID NO: 18) ATGGCGAGCTTCGCTTTAATAGTTATCCTTAGCGTAATTGGCTTTATATCGGCCTATCCT
AGTCCTGAAGGTTACAGTTCTGCCTTCAACTTTCCATTCGTAACCAAGGAGCAGTGGGGC GGCAGGGAGGCACGCACGTCGACGCCACTCAACCACCCAGTGCAGTTCGTGGTGATCCAC CACAGTTACATTCCCGGCGTGTGCCTCAGCCGGGACGAGTGCGCGCGCAGCATGCGCTCC ATGCAGAACTTCCACATGAACAGTAACGGGTGGAGTGATATTGGATACAACTTCGCTGTC GGCGGTGAAGGGTCGGTGTACGAGGGCCGCGGCTGGGACGCGGTCGGCGCACACGCAGCT GGCTATAACAGTAACAGTATCGGCATCGTGCTCATCGGCGATTTTGTTTCAAACCTCCCG CCGGCGGTGCAAATGCAAACCACACAAGAATTGATCGCAGCGGGCGTGCGACTCGGTTAC ATCAGGCCCAACTACATGCTCATCGGGCATCGTCAGGTCTCCGCCACTGAGTGCCCAGGA ACCAGACTCTTCAACGAAATCACCAACTGGAACAACTTCGTGAG
[0209] > M. sexta- Beta-1, 3-glucan-recognition protein 2 (MsPGRP2); AY135522.1 (SEQ ID NO: 19) .0 GCGTCTGTTTGTTCGCAACCATTGCGGGCTGCTTGGGCCAGCGAGGGGGTCCATACAAGG
TGCCTGATGCGAAACTCGAAGCTATCTACCCCAAAGGCTTGAGAGTCTCTGTGCCAGATG ATGGCTACTCCCTATTTGCCTTCCACGGCAAGCTCAATGAGGAGATGGAAGGTTTAGAGG CTGGCCATTGGTCCAGAGACATCACCAAAGCGAAGCAGGGCAGATGGATATTCAGAGATA GGAATGCTGAGCTGAAGCTTGGAGACAAAATTTACTTCTGGACTTACGTTATTAAGGATG GATTGGGATACAGGCAGGACAATGGAGAATGGACTGTTACAGAATTCGTCAATGAGAACG GTACAGTGGTGGACACTAGTACAGCGCCGCCACCAGTAGCACCCGCCGTTTCAGAGGAAG ATCAATCGCCAGGTCCTCAGTGGAGACCTTGCGAAAGATCCCTGACTGAGTCCTTGGCCC GCGAACGCGTTTGCAAAGGCAGCCTTGTCTTTAGCGAGGACTTTGATGGTTCCAGTTTGG CCGACTTGGGCAATTGGACCGCTGAAGTCAGATTCCCTGGCGAACCGGACTACCCGTACA ACTTGTACACTACGGACGGCACTGTGGGATTCGAAAGTGGGTCTCTGGTGGTGAGACCCG TCATGACCGAGTCCAAATACCACGAGGGCATCATATACGACCGCCTCGACCTTGAGAGAT GTACAGGACAGCTGGGTACGCTGGAATGCAGGCGAGAGAGCAGCGGCGGTCAGATTGTAC CACCTGTGATGACAGCTAAACTGGCCACTCGACGCAGCTTCGCGTTCAAGTTCGGCAGGA TCGATATAAAGGCGAAGATGCCGCGCGGGGACTGGTTGATACCAGAACTCAACCTCGAAC CTTTAGATAACATATACGGCAACCAGCGATACGCTTCGGGTCTCATGCGGGTCGCGTTCG TGAGAGGAAACGATGTATACGCCAAGAAGCTCTACGGAGGTCCGATAATGTCCGACGCGG ACCCGTTCAGGTCCATGCTGTTGAAGGACAAGCAAGGGTTGGCCAACTGGAATAATGATT ACCACGTCTACTCGCTGCTGTGGAAGCCTAACGGTTTAGAGCTGATGGTGGACGGTGAAG TGTACGGCACCATCGACGCTGGCGATGGCTTCTACCAGATTGCGAAGAACAACCTCGTGA GCCACGCCTCGCAGTGGCTCAAGGGCACCGTCATGGCGCCGTTTGATGAAAAGTTCTTCA TCACTCTGGGTCTTCGCGTGGCGGGTATCCACGACTTCACGGACGGTCCGGGCAAACCTT GGGAGAACAAGGGC
[0210] > M. sexta-Relish family protein 2A (MsREL2A); HM363513.1 (SEQ ID NO: 20) AGTGTGCGTCTACGAATGATTTGAAAATATGCCGCATAAGCCGTTGTTACGGTAGACCGA
GAGGCGGCGAAGATATCTTCATATTTGTCGAAAAGGTCAACAAGAAAAACATCCAAGTTC GGTTCTTTAGACTGGAAAACGGGGAGCGCACCTGGTCAGCGATGGCGAACTTTCTGCTAA GCGATGTTCACCACCAATACGCTATCGCTTTTAGAACGCCACCGTACGTCAATCACCAAA TTTCTGAAGACGTGCAAGTTTTTATAGAACTCGTACGCCCTTCAGACGGTAGGACGAGCG CTCCCATGGAGTTCACATACAAGGCTGAGCAAATCTATAAACAGAACAAGAAACGTAAAA CTACTTCGTCGTACTCGTCGCTCGACAGCTCCTCAGGTTCGGCCGGTTCAATTAAAAGCA
.0 TCAGCGAACTGCCCGCGCCCGTTGTTTTTGCTGAAAACGTAAGTTTTTTCTATGACACAT
TACTCATTCTTCAACCCATGACGAATCTATAA
[0211] > M. sexta-Dorsal (MsDor); HM363515.1 (SEQ ID NO: 21) ATGCTTGTGACGTTATGCGGCGGGAACTATAGTGGATTGTCGTTAACAAAAACTAATCAT
TATATGTCACCAAAATCATATGTGCCAGGAAATGGTTATGACGCCGCCGTAATCCTAGGT ACCACGGAGCAGAATGACAGCGAACCCTCAAACTTGAATATTAGTGATGTTTTTGAAGCC ATCACGCTCGCTGATCCGTCGTTCGGCGCGGGCGTGCCGTCGGTAGAGGAGACGATGGCG CACACGCAGCCCCAGCCGCTGCAAATGCCGTACGTGGTCGTCGTGCAGCAGCCCGCCAGC AAAGCGCTCAGATTTCGATATGAGTGCGAGGGCAGATCAGCCGGTTCGATTCCCGGCGCG TCGAGCACGCCCGAGAACCGAACCTTCCCCGCCATCAAGATAATCGGCTACACCGGCACC GTCTCCATCGTAGTGTCGTGTGTCACCAAAGATGAGCCTTGCAGGCCGCACCCACACAAC CTGGTCGGGCGCGACCACTGCGACCGCGGCGTGTTCTCCGTCCGCATCGAGATCACCGAC GAGAATAACGAATACCAGTTTCGGAACCTGGGCATACAGTGCGTCAAGCGGCGCGACATC GGCGAGGCGCTGCGGATCCGAGAGGACCTGCGCGTCGATCCGTTCAAAACCGGCTTCACC CACCGGAACCACCCGCAAGGCATCGATCTGAATGCAGTGCGGCTCGCGTTCCAAGTGTTC CTGCCGCACTCCAGCGGCAAGATGCGGCGCACGCTCGCGCCCGTCGTGTCCGACGTCATC TACGACAAG
[0212] > M. sexta-Spatzle (MsSPZ1A); GQ249944.1 (SEQ ID NO: 22) CGAACAACCTGACAGACGGATAGCGGGACGGTCAGCACAATACGAACATTTAAGAACAAA
CGAGAGGTCTCTCCCGGTCTACAGCGAGACCCAGAGGATACAAGCAGAAGAGAGAAGAAG ACACAGTTCGAGACTAGAAGAACCGAGACAACGTGCTGAGAATGGTTCATATAAGATATT GAATAACCCTCCGAAACCCTGTATTACTAATAGGAGAAGTCAAATTGATTCGTCGAATGA TAGGGTAGTGTTCCCCGGTCCGACTTCAGAAAGGTCGTACGTACCCGAAGTGCCAGAGGA ATGCAAGAAAATCGGCATATGCGACAGTATACCGAATTACCCAGAAGAACACGTAGCTAA TATTATATCTCGACTTGGAGACAAAGGAAAAGTATTACAAATAGACGAACTGGACGTATC AGACACTCCAGATATCGCCCAGAGGTTGGGTCCGCAGGAGGACAACATGGAACTATGTAG CTTTAGAGAAAAGATTTTTTACCCCAAGGCAGCGCCAGACAAAGATGGAAATTGGTTCTT CGTTGTGAATTCAAAAGAAAACCCAGTACAGGGTTATAAAGTTGAAATTTGCGACCGTCA GCAATTACCATGCGCGGAGTTCGCGAGCTTCCAACAGGGATATGAAGCGAGGTGCATCCA GAAATACGTTCGCCGGACCATGTTGGCGTTGGATCCCAAGGGTCAGATGACCGACATGCC CCTTAAAGTGCCCAGCTGTTGCT
~0
[0213] > M. sexta-Toll receptor (MsTOLL); EF442782.1 (SEQ ID NO: 23) CAAGTGGTCGTTGTGCACTGCACCGGACTCGCCGAGTTCCCGCGCCTGCCACTAACCACG
GACAAGTGGATCCTGCACCTGCCCCACAACAACATATCATACCTCGCCGCGGCCGACGTA TCGCCGAACATCGTGGAACTCGATCTAAGAAACAATTCAATCAAGAATATCGACGTACAG GCATCAGCAAAACTAGCCTTCGTCCGGCTGCAATTGGGTGGTAACCCGATCGAGTGTGAC TGCGAGGCGTTGAAGCTGCTGGCGCCGCTGCTCAAACCTGACTCAAAGCTGCTTGACAGA AAGGACGTGAAATGTGAGAACGACGCGCAGATTACCTTGGCGATGCTGAAATTATGTACT AAGTCATCCAATGGGCTGATGTACTTGCTGTTCCTGTTGCTGTTACTCGCATTTGTCGTA ACCGGACTGCTCGCACGAACCGCAATTCGCCTGCGCATCAAAATGATCCTCATGAGACTG GGTTGGATGTCGAGACTACTGGAGCCCGCGGACGACGATCGCCCGTACGACGCGTTTGTG TCTTTCGCACACGAGGATGAGGAGCTGGTGATGGAGCAGCTGGCGGCACGGCTCGAGAGC GGCTCGCGGCCGTACCGACTGTGTCTGCACTACCGCGATTGGGCGCCAGGCGAGTGGATC CCGGCGCAAATAGCGGCTTCGGTGCGGGCCTCTCGGCGCACGGTGGCGGTTGTGTCGGCG CACTACTTACAGTCGGGCTGGGCGCTTGCCGAGATCCGGGAGGCGACCGCAGCCTCGCTG CAGGAAGGCATGCCACGTCTCATCATCGTGCTGCTCGACGAGACCGACCGGTTGATGCTC GATATAGACCCTGAGTTGCACGCCTATGTGCGCAACAATACCTACGTGCG
[0214] > M. sexta-Scolexin A (MsSCA1); AF087004.1 (SEQ ID NO: 24) TCGAAGCTAAGGGGGCAAAGCAGCCAATTGATACGAGGGCAGTGAAGGAACGGTATCCAT
ACGCAGTTCGGAGTTTCGGAGGCTTCTGCGGAGGAACCATTATCAGTCCCACCTGGATCC TGACCGCCGGCCACTGCTCGATACTCTATGCGGGGAGCGGCCTACCGGCCGGCACCAACA TTACCGAGGTATCTAGCTTGTACCGCTTCCCCAAGCGGCTCGTCATACACCCGCTCTTCT CCATAGGACCCGTCTGGCTCAACGCTACGGAGTTCAACCTCAAACAGGCGGCTGCACGAT GGGACTTCTTGTTGATAGAACTGGAGGAACCGCTGCCGTTGGACGGCAAGATCCTGGCGG CTGCGAAGCTCGACGACCAGCCCGACCTCCCCGCAGGCCTCGACGTGGGCTATCCGAGCT ACAGCACCGACACCTACGAGGCTAAGATACAAAGCGAGATGCACGGAAAGAAGCTTTCGG TTCAATCTAACGAGGTGTGCTCGAAGCTAGAGCAGTTCAAGGCGGAGGACATGTTGTGCG CCAAGGGACGTCCACCGCGATACGACTTCGTCTGCTTCAGCGACAGTGGCAGTGGGCTAG TAGACAACAATGGTCGCCTAGTCGGCGTGGTGTCGTGGGCCGAGAACAACGCTTTCGAGT GCCGCAACGGCAACCTGGCGGTCTTCTCGCGAGTGTCCAGCGTACGCGAGTGGATCCGAC AAGTC
[0215] > M. sexta-Hemolymph proteinase 18 (MsHP18); AY672794.1 (SEQ ID NO: 25) GATGGTCAGAAAGCGTGGGACAAATGTCTGGAATACGTTGACAAGCTGTCGTACCCATGC
GCTTCAACCTACTCCCACTACCTCAGCTCCGTTTGGGAGAAAGATAAGGAGTGCAGTATG GTTCAGTTTGTTGGCGTGAGGCGATTCGCCTCGTATAACGGACAACCGGCGAAACGGAAC GAGTACCCTCACATGGCTCTGCTCGGCTACGGCGACGACCAGGAGACGGCGCAGTGGCTC TGCGGCGGCTCAGTGATCAGTGATCAATTCATCCTCACGGCTGCACACTGCATCTTTACA AATCTATTGGGTCCAGTACGTTTCGCAGCGCTAGGAATACTGCAGCGATCGGATCCAGTA GAGTTATGGCAAGTTTACAAGATCGGCGGCATAGTTCCCCATCCGCAGTATAAGTCACCT ATCAAGTACCACGACATTGCTCTCCTGAAGACTGAAAACAAAATAAAGTTTAATGAGAAC GTGCTGCCAGCGTGTTTGTTCATAGAGGGCAGAGTGGGTGGGAGTGAGCAGGCTAAAGCG ACCGGTTGGGGCGCGCTTGGACATAAACAGACGGCAGCTGACGTACTGCAAGTGGTTGAC CTTCAAAAGTTCAGTGACGAAGAGTGCGGAAGTACCTACCGTCCTTACCGGCATTTGCCT CAAGGCTACGACAGCGCCACGCAGATGTGCTACGGCGACAAGGGAAAACTGAATATGGAC ACCTGTGAGGGCGACAGCGGCGGTCCTCTACAGTTCCAAAACTCCTCGCTCCTCTGC
[0216] > M. sexta-Transferrin (MsTRN); M62802.1 (SEQ ID NO: 26) ATGGCTTTGAAACTTTTAACTTTGATAGCCCTGACTTGTGCGGCTGCGAATGCAGCTAAA
TCTTCATACAAACTATGCGTGCCAGCAGCATACATGAAGGACTGCGAGCAGATGCTTGAA GTACCCACGAAGTCTAAAGTGGCCTTGGAATGTGTACCGGCTAGAGACAGGGTGGAATGC CTCAGCTTTGTTCAGCAGCGACAGGCGGACTTCGTCCCCGTCGACCCTGAGGACATGTAC GTGGCCTCCAAGATCCCCAACCAGGACTTCGTCGTCTTCCAGGAGTACAGGACTGATGAA GAGCCTGATGCGCCATTCCGTTATGAAGCCGTTATTGTGGTTCACAAAGACCTACCCATC AACAACTTGGATCAGCTGAAGGGACTGAGGTCTTGCCACACCGGAGTCAATCGTAACGTC GGGTACAAGATCCCACTAACGATGTTGATGAAACGTGCCGTGTTCCCGAAAATGAACGAC CACAGCATTTCGCCGAAAGAGAACGAACTGAAAGCGCTATCGACGTTCTTCGCAAAGTCG TGCATCGTCGGCAAATGGTCGCCTGACCCCAAAACCAACTCGGCTTGGAAATCACAATAC AGCCATTTGTGTTCAATGTGCGAACACCCGGAGCGTTGTGACTATCCCGACAATTACAGC GGGTACGAGGGCGCGTTGAGATGCCTCGCCCACAACAACGGGGAGGTCGCGTTCACCAAA GTCATATTCACACGTAAATTCTTTGGGCTTCCAGTAGGTACCACTCCAGCGAGTCCATCA AACGAAAATCCCGAAGAGTTCAGATATCTCTGCGTGGACGGATCTAAAGCCCCCATCACT GGCAAGGCTTGTTCATGGGCTGCCAGACCTTGGCAAGGACTGATCGGTCACAATGACGTA
.0 CTTGCCAAACTCGCTCCGCTCAGAGAGAAGGTTAAGCAACTTGCTGATTCTGGTGCAGCT
GACAAACCGGAGTGGTTCACCAAAGTCCTTGGTCTATCAGAGAAGATCCACCATGTCGCT GACAATATCCCAATCAAGCCCATCGACTACCTGAACAAGGCTAACTACACGGAGGTCATT GAAAGAGGACATGGAGCTCCCGAGCTGGTCGTCAGGCTATGTGTGACGTCAAACGTGGCA TTATCTAAGTGCCGGGCTATGTCCGTGTTCGCATTCAGTAGAGACATCAGGCCGATCCTA GACTGTGTTCAAGAAAACAGCGAAGATGCCTGTCTTAAGAGCGTCCAAGACAACGGTTCA GATCTTGCCTCAGTAGACGATATGAGAGTAGCTGCAGC
[0217] > M. sexta-Arylphorin P subunit (MsARP); M28397.1 (SEQ ID NO: 27) CTGTCATAATCCTAGCGGGGTTGGTGGCCCTGGCCCTCGGCAGCGAAGTGCCTGTCAAGC
ACTCCTTCAAAGTTAAGGATGTTGATGCGGCTTTCGTCGAACGTCAAAAGAAGGTCTTAG ATCTTTTCCAAGATGTCGACCAAGTAAATCCTAACGATGAGTACTACAAGATTGGCAAGG AATACAACATCGAGGCTAACATCGACAATTACTCGAACAAGAAGGCCGTCGAAGAATTCT TGCAGTTATACAGGACAGGTTTCTTGCCTAAGTACTATGAATTTTCACCCTTCTATGACA GACTAAGGGACGAGGCCATTGGTGTTTTCCACCTCTTTTACTACGCTAAAGATTTTGATA CGTTCTACAAATCTGCCGCATGGGCGCGTGTGTACCTCAACGAAGGACAGTTCTTATACG CCTACTACATTGCTGTGATTCAGCGTAAAGATACTCAGGGCTTCGTTGTACCAGCACCGT ATGAAGTCTACCCTCAATTCTTCGCAAACTTGAACACTATGCTCAAAGTCTACCGTACCA AAATGCAGGATGGAGTTGTTAGTGCCGATTTAGCTGCACAACACGGCATCGTAAAGGAGA AAAACTACTACGTATACTATGCCAATTACTCCAACTCATTAGTGTACAACAACGAGGAAC AGAGACTGTCGTACTTCACTGAGGACATCGGCTTGAATTCGTACTACTACTACTTCCACT CTCACTTGCCTTTCTGGTGGAATTCTGAGAGATACGGAGCACTAAAATCGCGCCGTGGTG AAATCTACTATTACTTCTATCAGCAATTAATTGCACGTTATTACTTTGAACGTCTCTCGA ACGGCCTGGGTGACATTCCCGAATTCTCATGGTACTCACCAGTCAAGTCTGGCTACTATC CACTGATGTCTTCTTATTACTACCCCTTCGCTCAAAGGCCCAACTACTGGAACGTGCACA GCGAAGAAAACTACGAGAAAGTACGATTCTTGGACACGTATGAAATGTCATTCCTTCAGT TCCTCCAAAACGGACACTTCAAAGCGTTTGACCAGAAGATTGACTTCCACGATTTCAAAG CTATCAACTTTGTTGGAAACTACTGGCAAGATAATGCTGACCTGTACGGTGAGGAAGTTA CTAAGGACTACCAACGTTCATATGAAATTATAGCCCGCCAAGTGCTTGGTGCTGCACCTA AACCATTCGACAAGTACACATTCATGCCCAGCGCTTTAGACTTCTACCAGACGTCTCTGC GTGACCCAATGTTCTACCAACTTTACAACAGAATTCTGAAGTACATATATGAGTACAAGC AGTACCTGCAACCGTACTCTTCAGAAAAACTGGCATTCAAGGGTGTCAAGG
[0218] > M. sexta-Chymotrypsinogen-like protein 1 (MsCTL1); AM419170.1 (SEQ ID NO: 28) ATGTACGTGAAAGTAGCACTTCTGTTGGTAGCCCTCATTGCTGGGAGCTGGGCCTTCCCA
AAGCTCGAAGATGAGCAGGACATGTCCATCTTCTTCACGCAGCTCGATTCGAGCGCGCGT ATCGTGGGTGGTACCCAGGCCCCCAGCGGAAGTCACCCTCACATGGTGGCGATGACCACC GGTACCTTCATCAGGAGCTTCAGCTGTGGAGGCTCAGTTGTCGGTAGACGTTCCGTTCTG ACTGCGGCTCATTGCATCGCTGCTGTTTTCAGTTTCGGTTCCCTCGCCAGTACCCTCCGC TTGACGGTCGGCACCAACTTCTGGAACCAGGGAGGCACCATGTACACCGTCGCTCGCAAC ATAACCCACCCCCACTACGTCTCTGCGACCATCAAGAACGACATCGGTCTGTTCATCACT CACAACAACATCATCGACACGACTGTCGTCCGCAGCATCCCTCTTAACTTTGACTATGTG CCCGGTGGTGTTCTCACTAGAGTCGCCGGATGGGGCAGGATCAGGACCGGCGGTGCCATC TCTCCCTCTCTGCTGGAGATCATTGTGCCTACTATCAGTGGAAGCGCATGCGTAGCCAGT GCAATCCAAGCTGGCATCGATCTGAACATGAGACCACCTCCCGTCGAGCCTCACATCGAG CTGTGCACCTTCCACGGTCCTAACGTAGGCACTTGTAATGGTGACTCCGGCAGCGCTCTT GCCCGCCTAGACAACGGCCAGCAGATCGGTGTGGTATCGTGGGGCTTCCCGTGCGCACGC GGCGGTCCCGACATGTTCGTCAGGGTCAGCGCCTACCAATCCTGGCTGCAG
[0219] > M. sexta- Valine Rich Midgut Protein (MsVMP1); NCBI accession number not assigned as yet (SEQ ID NO: 29) ATCATTGACGGACCTTCCGTTGGACCNGCCATCATCGGCGCTGGAGACATCGCTGTCGGC
CCTGCTATCGTCGACTTCCCTTTCCCCGACGGCGGTGCCGTGTCTGCCCCCGTTGAGCCT TCCCCCATCGCCATCGGACCCGCTATCGTCGGTGAATCCCCTATCTCCGTCGGACCTGCC ATCGTTGAGGCCGGAGACATCGCTGTTGGACCCGCTATCATCGACTTCCCCCTTCCCGAC GGTGGCGCCGTGTCCGCCCCCGTTGAGGTTTCTCCCGTCGACTCCGTCGTCGTCGGCCCT GCCGCCGGCTCTCAGAGCTCTCCCCTCGTCCAGATCATCATCAACGTTAAGGCCCCCGCT GGTGCCGGCCCCGTTGTCGATGCCGTCGCTGACAAGCCCATGGACATCATTGATGTTATG CCCGTCGTCGACCCTGCTGATTTCGTGGACCTCACCCCCGTTGTAGAGCCTGTAGAAGTC GTCGACATTGTCGATGTCATGCCCGTGGTTGACCCCATCAACATCATCGATGTTATGCCT GTTGTTAAGCCCGTAAACCCCCTTGCCCGTT
[0220] > M. esculanta- Catalase 1 (MsCAT1) AF170272 (SEQ ID NO: 30) ATGGATCCTTGCAAGTTCCGTCCATCAAGCTCAAACAATACCCCCTTCTGGACCACCGAT
GCTGGTGCTCCAGTATGGAACAACAATTCCTCCATGACTGTTGGAACCAGAGGTCCAATC CTTTTTGGAGGACTATCATATGATAGAGAAACTTGCCAACTTTACCAGAGAGAGGATTCCA GAGCGTGTCGTCCATGCTAGGGGAATGAGTGCAAAGGGCTTCTTTGAAGTCACCCACGAT GTCTCTCACCTTACTTGTGCTGATTTCCTTCGAGCCCCTGGAGTTCAAACCCCTGTCATC GTCCGTTTCTCCACTGTTATCCACGAGCGTGGCAGCCCTGAAACACTCAGGGATCCTCGA GGTTTTGCGACTAAGTTCTACACCAGAGAGGGCAACTTTGATATTGTGGGAAACAACTTC CCTGTCTTCTTCATCCGTGATGGAATAAAATTCCCAGATGTGATACACGCTTTTAAGCCC AATCCCAAGTCTCACATCCAAGAATACTGGAGGATCTTTGACTTCTTATCACACCATCCT GAGAGCTTGAGCACCTTCGCCTGGTTCTTCGATGATGTTGGAATTCCCCAAGATTACAGA CACATGGAAGGTTTCGGTGTTCACACCTTTACTTTCATCAACAAGGCTGGAAAAGTAACC TACGTGAAATTTCACTGGAAACCCACTTGCGGGGTCAAGTGTTTGATGGA
[0221] > M. sexta-IMD (MsIMD); Msex2.05477-RA (SEQ ID NO: 31) ATGACTTCTTTGAAAAGCAAATTAGCAGAATTCTTGAAGGGGTTAAAATCAGATGCAACC
CCAAGCCCCGAGGCCATCGACAGAACCCAGGGTAAATCCACAAACTGAGCAATGAAAATG ACGCACCCTCGGATAGTGAACCTGAAGAAATCATTATAGAAGATATGACGATACGAAGAA AAAGAAAAAGGCAAGTTACAAGAAACCGTTCTTTAGCTCCAAACCTGGTGCATTCCCCGA AAAGAAAAAAGACAAGAATACCAAAGACGATTACAGCAACTTTGTGAATACTCAAGCCAC TGGTGATGTCATCAATATTGTAGGCTGTAACAATTTCCGCTGGGGTAATAATTATTATTT GGGAAATACCAAGAAACAGGCTCCTCCTAAGAAATATTTCCAAGAAGAGGAAGACAGTGA ACCAGAAGATGATATTCAGAAATGCAACTTAATCAAACTGCTATTTGAAGCTGAAAATAA GCCAGAACATGAGTACCTGGACTACATTTCCCAGAACATGAATGAAAAGTGGCACAGATT CTTTGTGAAACTAGGTTTTACACCAGGAAGGATCAAAACATCCATCATAGATAATGCCAG TTATGGTATTTCAGAGGCTCGATACGCATTGCTACTTGAGTGGGTGAATAAAAAACGGGA CTCTAATCTTGGACAGCTATCGAATTTATTGTGGAAACACGGCGAGAGGCGAATTGTCAA AGAATTAGCCATTATGTACTCTGCAAGCAAGGCCAAATCTGATGATGAATAG
[0222] > M. sexta-FADD (MsFADD); Msex2.03129-RB (SEQ ID NO: 32) ATGACTCTATCGGAATCAAAATTTAAACAATTAAAAGAACAAATTATTTTACATGCAAGT
GCAACTGAAAGACATGCTCAAATTTTGAACTCATTAAAGGATTTGTTTAAAGAAGATATT AATTCTGTTCGAAGATTTGAACAAATCAGCAATATAGCACAATTATTAAAAGTTTTAGAA ATAAGAGATGTGTTGTCAGAGGATGATGTTGCTCCTTTGAAAGATGTGGCACGCCAACTA CCAAATAGCTCAGAAATGCTGCGGAAAATTGCTGAATATGAAGAAAATCACAAGTGTGGA GAGTTTATTTCTGTTCCTAAGTCACCTCCAAAACAAAAAGAACATAGTCATTCTGGATGG
.0 GATATTAACAGCATACAGAATTCTGAATATTCTGTAAAAAAGGAAAGGATATTTGAAGTT
ATTTCTGAAGAAATTGGCACCCATTGGAGAAATCTAGCGAGATATTTGAAAACAAGGGAA TGTACAATAATTGAAATTGACAGTAAAAACATACCACTTTCAAGTAAAGCTATGGAGATT TTGAAATTGCATTGTAAAAAAGCAAATCCACAAAAGTGGTTTTTTGACTTGTGTAATGCA TTGGACAAAGTGCATAGGACTGACATAGTGTTATCACTGCAAGAAATTATGTCTATGAAT ATTTAG
[0223] > M. sexta-Dredd (MsDRD); Msex2.04297-RC (SEQ ID NO: 33) ATGTTGTCTTCCGATGCCAGAAGTTCCCTAAATTCCAAGAGTGATAATGAAACGTTTATA
TTAAACTTGGATTCGATTTCAAAGATTGAAAAACAATTACAAGATAATCCTTATGATATG ATCTCTTTAGTGTTCCTGTTGTACGACGTGCCGGATACGGCGCTGCAGAGATTGATGGTG TATCAACGAGTGACGAGTGACGTCAGTGGAACAAATATTAATTTGTTGCAAGAATGGTAC TGTCACGCCAGCAGTCGCCCTGACTGGCAACATGAATTATTAGAAGCATTAATGATATGC CAATTGAACTCCATTGTGAAGAGCCTTGGTTTCCATATACCCACTATGAGAGTATTTTAC CAATCAAATGATCCCTTCAGCAGCAAGTACATAAATCCCGTAAAGAAAGTCCTTTATCAT GCCTGTGAAAATATAAACTCAACTAATCTTTTGAAATTAAAAAAATCACTTCTTTCATAT GATATAAATGTAATGGAATATACAACTTGTGAGCTTATATTTTTAGAGCTCCTGTCCAAT AAATTTATTACTATTAATAATATTGGCCAAAAGATTAAAACAAACCAGAACTGCATATGT AATGTAGAAAACCTTGTGAAGATCTTAGACAACTTAAATGGATTGAAAAAAGTGGCTATG AATTTGAGATATTTTCAAAGTAAATTCAATGATGAAGAGGTGGATTCATTCGCGTCTGTC AATGGAAGCAGCTCACCATCACTTCCAGTGCCTCCTCTTGATGGTACAAAATTAAAGCAA GATGACAAGAATTATGGAGCTGAAGATTTCTCAGAGCTATTCGACATAATAAATAATATG CCAGAACCGCTTGAAAACAATTTTAAATCTGATACAATGTCAACTAAACAGAATAGATAT GAGATAAAAAATCCGGAACAACTAGGTGTTTGCATAGTTATAAACCAAGAGAATTTTTAT CCATCCAAAAATAGTATTGAAGACCACCAAATTGTTCCATTAGAAGAGAGAAAAGGATCT AGTGTGGATAAAAGGACTTTGGAAAGAACCATGACATCATTAAAATTTCAAGTTCACAGT TGCTCTGATTTAGATCATGATGAAATGATAGAATTCATCAAAAAGAAAATTAATAAACAT GTTACATCAAATGACAGTATTTTTATGTTGTGTATACTGTCACATGGTGTAAGGGACCAT GTGTATGCTGCTGACTCTGTGAAAATTAAAGTAGAGTCTATACAAAATCTGTTGGATTCA GATGAAATGAGCCACTTGAGAGGCATACCGAAGGTGTTTATTATACAAGCTTGCCAAGTT GAGGATACACCTCATCCTACATTTGCTGCTGACAATGTCCAAACAAATTACTACTTGGGA AAATTAGACTTCCTTATTTACTGGGCCACTGCACCTGAATATGAAGCCTTCAGACATGAG CAGACGGGGTCATTATTCATTCAAGCTCTTTGTAAACTGTTGCGTCAAAGAGCTAAACAT
.0 GATCACTTACATGAAATCTTTACACAAGTAAATAACAATGTTACCAATCTGTGCACTAAG
TTGCAGCGTGCACAAGTACCTCTCTTCAAAAGTACTCTGAGGAAAAATTTATATTTACAA GTGCCAGAATAA
[0224] > M. sexta-Relish F (MsReF); Msex2.08004-RE (SEQ ID NO: 34) ATGTCCTCTTGTCCAAGCGACTATGATCCCAGTGAATCGTCCAAATCTCCACAAAGTATT
TGGGAGTCAGGAGGATACAGTTCTCCGTCGCAACAAGTTCCTCAATTGACTTCTAACTTA ACAGAATTGTCTGTTGATCACAGCTATAGATACAATGGAAATGGACCATATCTACAGATC ACAGAGCAACCACAGAAATACTTTCGGTTCCGTTATGTTAGCGAGATGGTGGGAACACAT GGATGTTTGCTTGGCAAATCTTATACAACAAACAAAGTTAAAACTCATCCGACAGTTGAA CTCGTGAATTACACCGGTCGAGCCCTGATAAAGTGCCAACTGTCGCAAAACAAGAGCGAA GACGAACACCCGCACAAACTGCTCGATGAACAAGACAGAGACATGAGCCACCACGTTCCC GAGCACGGCAGTTATAGAGTGGTATTTGCCGGTATGGGTATAATTCATACTGCCAAAAAG GAAGTTGCAGGGTGGCTCTATAGAAAATATATACAGCAGAACAAGAATGAAAAGTTTAAT AAGAAAGAGCTCGAAGCGCATTGTGAGAGGATGTCCAAAGAGATCGATTTAAACATAGTT AGACTGAAGTTTAGCGCTCACGATATTGACACTGGCATTGAAATTTGCCGGCCAGTGTTC TCTGAACCCATTTATAATTTGAAGTGTGCGTCTACGAATGATTTGAAAATATGCCGCATA AGCCGTTGTTACGGTAGGCCGAGAGGCGGCGAAGATATCTTCATATTTGTCGAAAAGGTC AACAAGAAAAACATCCAAGTTCGGTTCTTTAGACTGGAAAACGGGGAGCGCACCTGGTCA GCGATGGCGAACTTTCTGCTAAGCGATGTTCACCACCAATACGCTATCGCTTTTAGAACG CCACCGTACGTCAATCACCAAATTTCTGAAGACGTGCAAGTTTTTATAGAACTCGTACGC CCTTCAGACGGTAGGACGAGCGCTCCCATGGAGTTCACATACAAGGCTGAGCAAATCTAT AAACAGAACAAGAAACGTAAAACTACTTCGTCGTACTCGTCGCTCGACAGCTCCTCAGGT TCGGCCGGTTCAATTAAAAGCATCAGCGAACTGCCCGCACCCGTTGTTTTTGCTGAAAAC TTACCTGAAAACAATGAGCGTATTATAGATATACCCGTACAACAGAATATGCTGTACCAG GTTCTGCCAAGTCAATGTGATTTAGCCGACGCGTTCATGGAGGTGGACAGCAAGGGAAGT CCGGCCAGCGTCGTGGACCCGATGTGGGCCGGCGCCGACGTCGGGCTGCAGATGGCGTCG AGCCTGCCCGCCATCAAACTCGGCTCCACCGAACTAGAGAGCTTGGCGCACCGCAGCCGG GACGGAGTGCCCATGGACAAAGAATTCCTTGACAATTATCTCAGCTCTCTCAACTCGCTT GGTGAACTTAACGACGATGATGAGATCAATCATATGCAATATGTGCGCTCGTTACAGATG GTCCACGCCGACTCGGCACGACGAAAGGCTGAACCGCAACCCAAAGACACTGTAACCAAA CCGACTCAATCTTCCCCGAAGGACACAGACTATGGCGCCCAGTCGGGACGCCCCACTGAA TATTCTGCTTATTATAAAATGGAAGACGGCGTGGAAGTCAAAAAACTGGTGAAAGAATTG
.0 TGTAGTATCATACAGAACAAAGCGGGATATAAAAAACAAGAAGTGAGAAACAAATTGGAG
AGGCTGTTCACCTACCGTCTGTCCAACGGAGATACATTCCTTCATATGACATTGTGCAGC AATCAAAGTAGTTTTGAATACATCGTCAAAATCATACATAGCGTGAAAATGACACATCTA TTGGACTATTGTAATAATAAACAGCAAACCATATTACACATGGCTATTGTAAACGACCTG CCTCGAATGGTCTCTTTACTTATTGCAAAAGGTTGCAATCCAATGAATAAAGACAGCGAG GGCGACAATGCGGTGCACTACGCTGTGAGGAGCGAATGTTGTTTAGAAGCATTATTGGAC GCCATCAAAAATAACAATGTTCGGTGCGATTTGAACGACTGCAATAACGAGAAGCAGACG GCGCTGCACCTGTCGACGAGCGGCGCGAGCGCGCGGCAGCTGGCGGCGCGCGGCGCCGAC CCGCGCGTGCGCGACGCGCAGGGCCGCACGCCGCTGCACCTCGCCGCCTACGACGACAAC TGCGACGTCGTCAGGGCGCTGCTCGAGTTCGTGTCGCCCTCGGAAATAGACGTTGTGGAC GGCTGTGGCAACACTGCGTTACAGATCGTCTGCGGCGGCTCCGTTAAGAAGAACACTTTG GAAATAGTAAAACTGTTGCTCCAAAAAAAGGCTGATCCGTTGAAGCAAGATGGGCACAAC ATATCGGCGTGGAAGATGGCGCGCGAACACTCCGAAATACGGGACGCGATGAAGGACTAT GTCCCCGCCGCCGCGTACGAGGAGGACACCAAGTCGGAAATGGACGATGAGTTCGAATCC GCTGATGAGGAGGACTACCGGATGGGTTCCGGATCGGGCGCTGTGAGTCTGCCGGAGCTG GGCGCATACGTGGAGGCGCTGAGCGCGGCGCTGGAGGCGCGCGGCGCGTGGCGCGCGCTC GCGCACCGCCTCGGCCTCGCCGCCGCGCTCGACTGGTGCGCGCGACAGCACGCGCCCGCG CGCACGCTGCTGCTGCACCTCAAGGAATGCAGAAACGACATATCCTCGAAAACGTTGGCT GTAATTCTAGAAGATATGGGAGAATTAGAGGCTGCTTCAATTATAAGGAGACACATAGAG TGA
[0225] > M. sexta-Cdc42 (MsCdc42); Msex2.04668-RA (SEQ ID NO: 35) ATGCAGGCGATCAAGTGTGTCGTCGTCGGAGACGGTGCCGTCGGTAAAACATGTCTGCTC
ATCAGCTACACGACAAATGCCTTCCCCGGAGAATACATACCTACAGTATTCGACAATTAT TCAGCGAATGTGATGGTGGACGGGAAGCCGATCAACCTGGGCCTGTGGGATACGGCGGGG CAGGAGGACTACGACCGGCTGCGGCCGCTGTCCTACCCACAGACCGACGTGTTCCTTATA TGCTTCTCGCTCGTCAACCCGGCTTCGTTCGAGAACGTTCGGGCTAAGTGGTACCCAGAA GTGCGGCATCACTGCCCGTCGACGCCCATCATCCTCGTCGGTACCAAGCTGGACTTGCGC GAAGACAAAGACACCATAGAGAAACTTAAGGACAAGAAGCTCGCGCCTATCACTTACACA CAGGGTCTGGGCATGTCGAAGGAGATCAACGCGGTGAAGTACCTCGAGTGCTCTGCGCTG ACGCAGAAGGGTCTGAAGACGGTGTTCGACGAGGCCATCCGCGCCGTGCTGTGTCCCGTG CCGCCCCCCAAACAGAGCCGGAAGTGCACGCTGCTGTAA
[0226] > M. sexta-DSori (MsDSorl); Msex2.00725-RB (SEQ ID NO: 36) ATGAGTAAGATGTCAAAGAATAAATTAAACTTGACCCTGCCACCAGGGTCAATAGACACA
GCACCAGCCATCACACCATCCAATATGACACCACAGCTGAAGTCCGCAACAGCTACGGAG CGTCAGGGCTTGGCTGGTAAATCGAAAACCAGCATCGAAGCCCTGACAGAGAGGTTGGAG CAAATCGAGATGGACGACACACAGAGACGGAGAATAGAAGTGTTTCTGTGTCAGAAGGAG AAGATCGGGGAGCTTAGTGATGATGATTTTGAAAAGCTTGGAGAGTTAGGCCAAGGCAAC GGTGGCGTTGTAATGAAAGTCCGTCACAAGTCAACCGGTCTGATAATGGCGCGAAAGTTA ATCCATCTGGAAGTCAAGCCGGCAATAAAGAAGCAAATCATCAGGGAGTTGAAAGTCTTA CACGAATGTAACTTTGCGCATATCGTCGGCTTCTACGGGGCCTTTTATAGCGACGGCGAG ATCTCGATTTGTATGGAGTACATGGACGGTGGGTCCTTAGACCTTATACTGAAGAAGGCC GGCAAGATTCCTGAATCTATTCTAGGAACAATAACATCCGCCGTGCTGAAAGGTCTGAGC TACCTCCGGGACAAGCACGCCATCATGCATCGCGACGTGAAACCATCAAACATTCTGGTG AACAGCAACGGCGAGATCAAGATATGCGATTTCGGCGTGTCCGGTCAGCTGATCGATTCC ATGGCCAACTCTTTTGTCGGCACTAGGAGTTATATGTCTCCCGAACGTCTCCAAGGCACG CACTATTCAGTCCAATCTGACATATGGTCGCTCGGGCTGTCGTTGGTAGAGATGGCAATC GGAATGTATCCCATACCGCCGCCGGACGCGAAGACCCTGGCTGCCATCTTCGGTGGACAG AATGAAGATCATTCTCCTGGTCAGGCGCCGAACTCGCCCCGTCCGATGGCCATATTCGAG TTGCTGGACTACATCGTGAACGAGCCGCCGCCGAAGCTGCCCGCCGGAATATTCTCCGAC GAGTTCAAGGACTTCGTCGACCGCTGCTTGAAGAAGAATCCAGACGAACGAGCCGACTTG AAGACTTTGATGAATCACGAATGGATACGCAAAGCGGACGCAGAAAAGGTGGACATAGCG GGGTGGGTGTGCAAGACAATGGACCTCATGCCTTCCACTCCAAACTCTAACGTGTCTCCT TTTTCTTCATAA
[0227] > M. sexta-FOS (MsFOS); Msex2.09858-RA (SEQ ID NO: 37) ATGCAGAACATCGATCCTCTGGAGATCGCCAACTTCCTCGCCACGGAGCTGTGGTGCCAG
CAGTTGGCGAACCTCGAGGGCCTCCAGTCGGGGGTCCCGACTCGCACAACGGCCACCATC ACGCCGACGCAACTGCGCAACTTCGAGCAGACTTACATCGAGCTGACCAACTGCCGCAGC GAGCCCACCACGCACGCCGGCTTCGTGCCGCCGTCAGTCACGCACGCCAACAACTACGGC ATCCTGAATCCGACGGCGTATTGTGATTCGGGCCCGACGACGGCGCTGCACGTGTCGCCG GGCCCGCTCTCCGCCAGCGGCGACAGCAGCAGCAGCCCCGGCCTGCCCACGCCCAAGCGG CGCAACATGGGCGGCCGCCGGCCCAACAAGGCGCCGCAAGAACTCACGCCCGAGGAAGAA GAGCGCAGGAAGATCCGCCGCGAGCGAAACAAAATGGCCGCCGCCCGTTGTCGCAAGCGC AGACTCGACCACACCAACGAGCTGCAAGAGGAAACCGACAAATTAGAGGAAAAGAAGCAT
.0 GCGCTCCAGGAGGAAATCCGCAAACTGAACGCTGACCGGGAGCAGCTGCAAGTGATCCTT
CAGAACCATATGGTATCATGCCGGCTCAACAAGAGATCCATCAGCCCGCCCGATGTGAAG CCCTTCCAGGACCCGTACGCCTACCCTGAGATACCCGAGGATGGCGTCCGTGTCAAGGTG GAAGTGGTCGACCCCTCAGTAGACACGGTCTTAGTGTTGGACAATATTTACTCAACGCCG CCGACTGACAAAAAAATTATGCTGTCGTCGGCCAACCCAGCCGTGGTGACGAGTGGGTCG CCCGCCGCCCTGGAGACCCCGCCGGCGATAGTGCGTCCCAACAGACCCAACTCCCTCCAG GTGCCGCTCAGCCTCACACCAGCACAGTTACACAACAACAAGGCGCTGGGAAACAACAAA ATAGCCGGCATAGAGATAAGCACGCCGAGCAACGGCATCCCGTTCAACTTCGAGAGCCTG ATGGAGGGCGGCACGGGGCTGACGCCGGTGCACGGCCACGCGCTGCCGCTGCCGCACCCG TGCGCGCAGCAGCAGCGCGCCGCGCCCGACGCCTCGCCCGCCGAGCCGGCGCCGAGCTCG CTGGTGAGCCTCTGA
[0228] > M. sexta-Jra (MsJra); Msex2.12422-RB (SEQ ID NO: 38) ATGGTTCGGCACTCCGGCCACGGCATGGAGACCACTTTCTATGACGAGCAGTATCCCATC
AGCGGCCCCGTGGAGAACCTGAAGCGGCCCCTCACGTTAGACGTGGGGCGCGGCGTGAAG CGCGCCAGGCTCGGCGGGGCACCCGTACTCTCATCTCCAGACCTACAGATGCTGAAGCTC GGCTCGCCAGAGCTCGAGAAACTGATCATCCAGAACGGCTTGGTGACGACGGCCACCCCC ACTCCAGGTGCGCCGGTGCTGTTCCCCGCGGTCGCGCCTACCGAAGAGCAAGAGATGTAC GCGCGGCCATTCGTCGAGGCGCTAGACAAGCTGCACCACGGCGAGGTGACCCCGATCGGG CGGCGAGTGTACGCCGACCTGGACCGGCCGCTCGAGCGGTACCCCACGCCCGTGGTGAAG GACGAGCCGCAGACGGTGCCCAGCGCCGCCAGCTCGCCTCCACTTTCCCCTATTGACATG GACACGCAGGAGCGGATCAAACTGGAGCGCAAGCGACAGAGGAATCGAGTGGCCGCTTCC AAATGCAGGCGGCGCAAGCTCGAACGCATCTCCAAGCTGGAGGACAAGGTGAAGATCCTG AAGGGCGAGAACGCGGAGCTGGCGCAAATGGTGGTGAAGCTAAAAGAGCACGTACACCGA CTGAAGGAGCAAGTGCTGGAGCACGCCAACGGCGGCTGCCACATCGAGTCGCACTTCTGA
[0229] > M. sexta-Caudal (MsCAD1); Msex2.04570-RA (SEQ ID NO: 39) ATGGTGAATTACGTTAATCCCCTCGCCATGTACCAAGGCAAGGGCGGGCAATACGGCGGC
GGGTGGTACGGCTGGCAGCATCAGAACTTTGAAGAACAACAATGGTGTGCTTGGAACGGT GCGCCGGCGGGTGGCGAGTGGGCGCCAGATCCACATCATTTTCCCAAAAGAGAACCTGGA GAGAGAGAAATAGCAGACATGCCATCACCGGCACGAGGAGACTTGGCAAGTCCAGCAGAA
.0 GGTTCGCCGAGCTCGGGGTCGAGGCCGTCGCAGCCGCCAGCACCGCCGCGTTCCCCATAC
GAATGGATGAAAAAACCCAACTACCAGACACAACCTAACCCAGGTAAAACGCGCACAAAA GACAAATACAGGGTGGTGTACAGTGATCATCAGAGGCTGGAGTTAGAGAAGGAGTTCCAC TACAGCAGGTACATCACTATTCGTCGCAAGGCAGAACTCGCCGTCAGCCTTGGCCTTTCC GAACGACAGGTCAAAATTTGGTTTCAAAATAGACGAGCCAAGGAAAGGAAACAGGTGAAG AAACGCGAAGAAGTGGTGATGAAAGAGAAAGGCGACCATGCATCTCTACAGCACGCGCAG CTGCACCATGCCACCATGCTGCATCACCAGCAGATGATGAACGGCATGATGCACCACCAC CACTACCACCAAGGCGTGTTGCAAGGCGTGCCGGAGCCGCTGGTGCCGGGCGTGCCGCCC GTGCCGCTGCTGTGA
[0230] > M. sexta-Atg8 (MsAtg8); Msex2.12227-RA (SEQ ID NO: 40) ATGAAATTCCAATACAAAGAAGAACATTCTTTCGAAAAGAGGAAGACTGAAGGCGAAAAG
ATTCGCAGGAAATACCCGGATCGCGTTCCAGTAATTGTAGAGAAAGCCCCGAAGGCTAGA TTGGGAGACCTCGATAAGAAGAAGTATTTGGTGCCGTCTGATTTGACTGTCGGACAATTT TATTTCCTAATTAGGAAACGCATCCATCTTCGGCCTGAGGACGCATTGTTCTTTTTCGTG AACAATGTTATTCCACCAACATCCGCCACCATGGGCTCTCTGTACCAGGAACATCATGAC GAAGATTTCTTCCTCTACATTGCATTTTCTGATGAAAATGTTTATGGATTTTAA
[0231] > M. sexta-Atgl3 (MsAtgl3); Msex2.06273-RA (SEQ ID NO: 41) ATGGCACCCGAAGTGGCTTTTTCTAACATAAATGACAAGAATGAATTTACTAAATTCACA
AAATTCTTAGCTTATAAAGGCGTTCAAGTTATTGTAGAATCGAGGAAGGGTGTAAAAATA GATCCTAATAGTAAACCAAGATCATCAGACACTGATTGGTTCAATCTTCAAATACCAGAC TCTCCAGAGGTTAACCAGGCAACTAAGAATGCATTGCCTTCAGACAAGGTGTTAGAGATT ATCAAAGCCCAACTGCACGTGGAAATATCAGTACAAACCGAGGATGGCGATGAAATGGTT TTGGAGTTGTGGACCCTTGAACTTGACGAAACTCAGTTTGACACTTCTGTTAAAGCCACG AACACAGTTTACTTCAGAATGGGAATATTACTTAAGTCGCTTATAACTATAACAAGAATT ACTCCAGCGTACCACTTGTCGAGGAAGCAAAGAACAGAGTCGTTCACAATATTCTACAGA GTATACAATGGCGAACCGAAAGTAAAAGCGCTTGGAGAATCAGTGAAGAAAATTCAAGCT GGAATGCTCAAAACTCCACTTGGAGGGATAATATTCTCCGTGGCCTACCGCACAAACTTC TCCATTTCGCCAAACAGATCGGAGAAGGACAAAACATTGCTTTTGAAGAGTGATCATTTC GAACTGAGTCCAAAACATGTGATATTTGAATCCAAGAAAAAGAAAGACGAGAAAAAAGAA TACAAGCCTCTGAAACCTGTTGATTTGAATAAGCCACTTCGATTAGCCGCATTTGTAGAC GAGGATGTTGTAAAAAAGGCCTTTGATGACTTCATGGAGAAGATGCCGATTCCCAAATAC
.0 CGAGTGATACGTAGGGAGAGAAGTCCAGAGAGCGACAAGCCTATAGTATCCAAAAGTACT
CAGGATTTGGACGCTTCTGTGACGTCAAAGAGTCCGACGTCAATGGAAATGCCACCTAAA AAGTTCACAGGCTTTCGCAATGAGAACGAACCTCCATTAAAACTTCTCCATTTCCCATTC GCTGATAACCATCCGATAAGAGAACTGGCCGAGTTTTACAAAGAGTTCTTCAACGCCCCC CATTTGAAGTTAGCTGATGACGTTAGCCTGAAATCGGGCAGTGCTGAATCGGTAAAAGAG GTGATAGAGATTACAACTGAAGATTTGTCGAAAGATCTCGAATTGTATGAGAACTCGGTG TTGGAGTTTGATGAGCTCTTGGCAGATATGTGCCGATCGGCTGAGTGGAGTGGGAACTAG
[0232] > M. sexta-IAP1 (MsIAP1); Msex2.05607-RA (SEQ ID NO: 42) ATGGCATCAGCTGGCGCCGTACCGAATATTCTAGTGTCGTCGTCGCTTTTCAAGACCACT
CCTCGGGATCCTAAAATTCGTCGAAAAACAAGCCCTTTGAAGTTACCACCTTGTGACACA CATATAGGCTCGCCTCCACCATCATCCTCTCCGACCTCCTCGTCAACTGATAAAACCGAT
AATCATGATACTTTTGGTTTCCTTCCTGACATGCGtgatatgcgCCGCGAAGAAGAACGA
CTGAAAACTTTCGATAAGTGGCCTAGCACGTACGTGACACCTGAAGAGTTAGCTCGTAAT GGTTTCTACTACCTTGGGCGCGGTGATGAAGTTCGCTGTGCATTCTGTAAAGTAGAAATT ATGCGATGGGCGGAAGGAGACGACCCTGCGAAAGATCACAAACGATGGGCGCCCCAATGT CCGTTTTTACGTAATCTTTCGAACGGTACTAATGGCAGCGGAGAAGGTAGTTCGGAAGGT CGCGACGAGTGTGGTGCGCGAGCAGCGACGCGGGAGCCTGTGCGTATGCCCGGACCTGTG CATCCCCGCTATGCAACAGAGTTATCGCGTCTCGCCAGTTTCAAGGATTGGCCGCGTTGC ATGCGCCAAAAACCAGAGGAACTCGCGGAAGCAGGATTTTTCTACACCGGCCAAGGGGAT AAAACGAAATGTTTCTATTGCGATGGAGGTCTCAAAGATTGGGAAAACGACGATGTTCCA TGGGAACAGCATGCCCGTTGGTTCGATCGTTGTGCGTATGTGCAACTTGTTAAAGGCCGT GATTATGTCCAGAGGGTATTATCGGAAGCGTGTGTTATACGTGCCACCGACGAAAAGCCT GCGCCTGCACCTCAACCGTCCCAACCAAATGTCTCAGTTGTATCAGAAGAAAAACCTGTC GAAGAGGCTAAGATATGCAAAATATGTTATTCCGAAGAACGAAACATATGCTTTGTGCCG TGCGGTCATGTGGTCGCTTGTGCAAAGTGTGCCTTATCTACTGATAAATGCCCAATGTGC CGGAGGGCTTTCACAACTGCACTGCGACTCTATTTTTCGTGA
[0233] > M.sexta-Chitin synthase 2 (MSCHS2); AY821560.1 (SEQ ID NO: 43) ATGGCCGCAACTACACCAGGTTTTAAGAAGTTAGCAGACGATTCTGAGGATTCAGATACA
GAATACACCCCGCTGTATGATGACGGTGATGAAATAGATCAAAGAACTGCACAAGAAACA AAAGGATGGAATCTATTTCGAGAGATTCCGGTGAAAAAGGAGAGTGGATCTATGGCCACA AAAAATTGGATAGAAACAAGCGTAAAAATCATAAAAGTGCTTGCCTACATATTGGTTTTT
.0 TGTGCTGTACTGGGTTCCGCAGTCATAGCTAAAGGAACTCTTCTATTTATTACGTCACAA
CTGAAGAAAGACAGACAAATTACTCACTGCAATAGACGACTTGCTTTAGACCAACAGTTC ATAACGGTACACAGTTTAGAAGAAAGAATAACATGGCTATGGGCAGCACTTATTGTATTC GGTGTGCCGGAGTTAGGGGTGTTTTTGAGATCCGTCAGGATATGCTTCTTCAAAACTGCC AAGAAACCAACCAAAACACAGTTTATTATTGCTTTCATAACAGAGACACTACAAGCAATA GGAATAGCAGCACTTGTATTAATAATTCTACCAGAATTAGACGCTGTGAAAGGAGCCATG TTGATGAACGCCACGTGCGCTATCCCTGCATTGCTAAACATTTTCACGAGAGACCGAATG GATTCTAAGTTTTCTATAAAATTGATATTGGATGTATTGGCGATATCGGCACAAGCCACG GCGTTTGTTGTTTGGCCTCTTATGGAAAGAACGCCAGTTCTATGGACCATACCAGTTGCA TGTGTGTTAGTGTCTCTAGGCTTCTGGGAGAATTTTGTTGACACCTACAATAAAAGTTAT GTTTTTACGGTGCTGCAGGAACTACGCGACAACCTCAAGAGGACTCGGTACTACACTCAG CGGGTGCTATCTGTTTGGAAGATTATAGTGTTTATGGCATGCATTTTAATATCGCTGCAT ATGCAAAATGACAATCCGTTTACCTTTTTCACTCACGCCAGCAAAGCCTTTGGAGAGAGA CAGTATGTCGTTAACGAGGTTCTAATAGTAGTCCGAGATGACGAAACCATAGGCTATGAC GTCACCGGAGGTATATTCGAATTGGACGCGATATGGACCTCAGCATTGTGGGTCGCATTA ATTCAAGTGGGAGCAGCCTACTTCTGTTTCGGAAGTGGCAAGTTTGCTTGCAAAATTCTT ATACAAAATTTTAGTTTCACTTTAGCATTGACTCTCGTCGGGCCCGTGGCAATCAACCTC CTTATTGCTTTCTGCGGAATGAGAAATGCAGACCCTTGCGCTTTCCATAGAACTATACCT GACAATTTGTTTTACGAGATACCACCTGTGTACTTCTTGCGGGAGTACGTGGGCCACGAG ATGGCGTGGGTGTGGTTATTGTGGCTCATATCTCAAGCGTGGATCGTGTTTCACACGTGG CAGCCGCGATGCGAGCGCCTCTCCGCAACTGACAAACTGTTTGCCAAACCGTGGTACATC GGACCGCTAATCGACCAATCGTTGCTGCTAAACAGGACTAAGGATTTGGATAATGATTGC CAGGTTGAGGATTTGAAGGGTCTTGGCGACGATTCGTCGGTTGGAAGCGATCTTGCCATC GTAAAAGATATCAAACCGTTCGATTCGATAACCAGAATACAAGTGTGTGCGACAATGTGG CACGAGACCAATGAGGAGATGATCGAGTTCTTGAAGTCGATATTCCGCCTCGACGAGGAC CAGAGCGCGCGCCGCGTGGCGCAGAAGTACCTCGGCATCGTCGACCCCGACTACTACGAA CTCGAGTGTCATATTTTCATGGATGACGCTTTTGAAATATCCGATCACAGTGCCGAAGAC TCGCAGGTGAATCGCTTCGTGAAGTGCCTAGTGGATGCGGTGGACGAAGCGGCTTCCGAA GTGCATCTGACTAACGTGAGGTTAAGGCCCCCCAAGAAATACCCCACCCCGTACGGCGGA AAACTAATTTGGACCATGCCAGGGAAAAATAAGTTGATTTGCCATTTGAAAGATAAATCC AAGATTCGGCACAGAAAGAGATGGTCGCAGGTTATGTACATGTACTATTTCCTGGGGCAT CGCTTGATGGACCTGCCAATATCCGTGGATCGTAAGGAAGTGATTGCCGAGAACACGTAC CTATTAGCTTTGGATGGAGACATCGACTTCAAGCCGAGCGCTGTGACGTTGCTGGTCGAT
.0 CTTATGAAGAAGGATAAGAACTTGGGCGCCGCTTGCGGTCGCATTCATCCTGTCGGCTCT
GGTTTCATGGCCTGGTACCAGATGTTCGAGTATGCTATTGGTCATTGGCTGCAAAAGGCG ACTGAACACATGATCGGCTGCGTACTCTGTAGTCCGGGATGCTTCTCCCTCTTCAGAGGA AAGGCGCTTATGGACGACAACGTCATGAAGAAATACACACTCACTTCTAACGAGGCTCGA CATTACGTGCAATACGATCAAGGCGAGGACCGTTGGCTGTGTACGTTGCTGCTGCAGCGC GGGTACCGCGTGGAGTACTCGGCGGCGTCGGACGCCTACACGCACTGCCCCGAGCGGTTC GACGAGTTCTTCAACCAGCGCCGCCGCTGGGTGCCCTCCACCATGGCCAACATATTCGAT CTGCTCGCGGACTCCAAACGCACCGTGCAAGTCAACGACAACATTTCCACTCTGTATATC GTCTATCAGTGCATGCTTATGATGGGTACGATTTTGGGTCCGGGAACAATCTTCCTGATG ATGATTGGCGCAATAAACGCTATAACAGGCATGAGCAATATGCACGCACTTCTCTTTAAC CTGGTGCCCGTGCTTACGTTTTTGGTTGTCTGTATGACATGCAAGTCCGAGACTCAGTTG ATGCTCGCGAACCTCATTACCTGCTTTTATGCGATGGTAATGATGTTTGTGATCGTCAGT ATCGTCCTACAAATATCACAAGATGGTTGGCTAGCGCCATCTAGTATGTTCACTGCGGCA ACATTTGGAATATTCTTCGTAACAGCGGCTTTGCATCCACAAGAAATAATATGTTTGTTG TACATTTCCATATATTACATCACAATTCCGAGCATGTATATGTTGTTGATTATCTACTCC CTATGCAATTTGAACAACGTCTCGTGGGGAACTCGAGAGGTGGCTCAGAAGAAGACTGCA AAGGAAATGGAAATGGATAAGAAAGCAGCGGAAGAAGCAAAGAAGAAGATGGATAATCAA AGCATAATGAAATGGTTCGGCAAGTCGGACGAGACGAGCGGCTCGCTGGAGTTCAGCGTG GCGGGACTGTTCCGCTGCATGTGCTGCACCAACCCTAAGGACCACAAGGACGACTTGCAT CTCTTGCAGATCGCCAACTCCATCGAGAAGATCGAAAAGAGATTGTCGGCACTTGGCGCG GAGGAGTCCGAGCCGGCGCAGGCGCAGACGCGGCGCCGCTCGTCGCTGGGGCTGCGGCGC GACTCGCTCGCCACCATGCCCGAGTACGCCGACAGCGAGCTGTCCGGAGACATTCCTCGC GAAGAAAGAGACGATCTTATAAACCCCTATTGGGTGGAGGATCCCAATCTGCAGAAAGGT GAAGTAGACTTCTTGACGACGGCTGAAATCGAATTCTGGAAGGACTTGATTGATGTTTAT TTGAGGCCTATCGATGAAAACAAGGAAGAGCAGGAACGTATCAAAACCGATCTAAAGAAC TTGCGTGACACGATGGTGTTCGCGTTCGCCATGTTGAACTCGTTATTCGTGTTGGTGATA TTCCTGCTGCAGTTCAACCAGGACCAGTTGCACATAAAGTGGCCGTTCGGGCAGGATGTC GCGCTTTCTTATGACAAGGAAAGGAATGTTGTATTGGTGGAGCAGGAATTCCTTATGTTG GAGCCTATAGGTTCCCTGTTCCTCGTGTTCTTCGGGTTTGTAATGTTGATACAGTTCGTG GCGATGTTGTCCCATCGTTCGTATACTATTACGCATTTGCTCTCCACAACAGAGCTTCAC TGGTATTTCAGCAGACGTCCGGACCAGATGTCAGATGAAAACCTCTTGGAAAGGAAGGTG GTAGAAATAGCGAGAGAGTTGCAGAAGTTAAACACGGACGACCTAGACCGCCGCGCGGTC GAAACTAACGACGTGTCGCGGCGAAAGACTCTACACAACCTAGAGAAGGCGCGAGACACC
.0 AAGCACAGCGTGATGAACCTTGACGCTAACTTCAAGAGGCGACTGACTATACTACAGAGC
GGTGATCCTAACGTGATATCTCGGCTGTCATCGTTGGGCGGCGATGAGGTTACTCGTCGC GCCACGATACGCGCATTAAAGACGAGGAGGGACTCACTGCTCGCTGAAAAACGACGCTCC CAGCTGCAAGCGGCGGGCGACGCTACAGGCTACATGTATAACCTGTCAGGCACTGCGGTG AACGACATGAGCGGCCGAGCTTCGACGGCCAGCGCCTACATTAATAAAGGATACGAACCC GCTTTCGATAGCGACGACGACGAACCACCGCGTCCGCGCAGGAGCACTGTACGCTTCAGA GAAAACTACACGTAA
[0234] > M.sexta-Beta-fructofuranosidase 1 (MsSucl); GQ293363.1 (SEQ ID NO: 44) ATGTACATTAAAACAGCAACATTTTTGCTGTGCGTTTTCCTTGGTAGTGTATCGTCATGT
TGCGTTAATGGGCGGTACTACCCGAGGTACCATTTGTCGCCACCGCATGGCTGGATGAAC GACCCCAACGGATTCTGCTACTTCAAAGGTGAATACCATATGTTTTACCAGTACAATCCC ATGTCAAGTTTGGAGGCTGGCATAGCTCATTGGGGTCATGCGAAAAGTAAAGATTTGTGC CATTGGAAACACTTAGACCTCGCCATCTATCCTGATCAGTGGTACGATCAAACGGGAGTA TTTTCTGGAAGTGCGCTAGTAGAGAATGACGTCATGTACCTTTATTATACTGGAAATGTA AATCTTACTGATGAAATGCCATTTGAGGGACAATTCCAAGCTCTTGGTATCAGTACTGAC GGTGTCCACGTAGAAAAGTATAAAGACAATCCAATAATGTACACGCCAAACCATCAACCT CACATCCGAGACCCAAAAGTTTGGGAACACGACGGCTCTTATTATATGGTCTTAGGAAAC GCATATGATGATTATACAAAGGGCCAAATAGTTATGTACGAATCATCAGACAAGATCAAC TGGCAAGAAGTAACTATACTATATAAATCAAATGGATCTTTCGGTTACATGTGGGAGTGT CCAGATTTATTCGAAATAGACGGCAAGTTTGTACTTCTGTTCTCTCCTCAAGGCGTGAAG TCTGTGGGCGATATGTACCAGAATCTGTATCAAGCAGGATACATCGTCGGAGAATTCGAT TACGATACTCATTCATTCACAATACTAACCGAATTCAGAGAATTGGATCACGGTCATGAT TTTTACGCTACACAAACAATGAAAGATCCTAGTGGAAGAAGAATAGTCGTTGCTTGGGCA AGTACTTGGGAGTATGCTTATCCTGAACGAGCAGATGGTTGGGCTGGCATGCTCACACTA CCTAGAACTTTAACTTTGACAAAAGATTTAAGACTAATCCAAACTCCAATTAGAGAGATC GATCAAGTTTTTAGAAGAAGACTATATTCAGGAAAAGCCTCAGCAGGCAAAACTGTCGCT TTACCAGACAAAGCAGGGAAAGTAGAACTGAAATGGGATACACCAAGAAATATAAAGGTA GTTATAGAATCTCAAAATGAGTGCCAAAACGTAGTAATCAGTTATGATCACGAGGATGGT ACTATTACTTTGGACAGAGGAGGCGATGACGCAATCCGCCGCACTCACTGGGATCCTCGA GGTCACCTCAAATGGACCATTTTTATTGACGCAAGCTCCATAGAACTCTCTTGTGGTGAT GGAGAAGTATGGTTTACAAGCAGATTTTTCCCTGAAGGAGTCGTATCTGTTCGCCTTGGA
.0 GAAGATACTTGTGTTGATAAGTTTACCGTGCATTCTATTCGCCGTACTACTCCAGACCCC
GAGGCTCATTGTCGTTGTGAATCAGAAGAATAA
[0235] > M. sexta-Hemolin (MsHEM); M64346.1-UTRs
5'UTR (SEQ ID NO: 45) TGTTACATTAATTATAAAAAAAAATACAAAAAAGAAATATATAAGTAAGTACTAATAATA
TGGTCTAGTTTATTATAAGTCTTAGAGACTAACGATAAATAATAAATTTCTCATGAACCA CCTATTTATTTATTTGAAAAGCATTTTAAATATATTATAGATTTAAACGTAACGAAGTCT TTAAACTCGCGGGTAAAAACGCATACTTACTTGACTCTACATCGGCGACACACGAAAAAC TTATTCGTACTGTTTTTTCATTGAATATGTTATGAAAAAGAAAACTATATGTAATCGCAG TCACATATATAAAACTTTATAAAAAAGTTAACAGAGATAAATAACTTGTTTGGTGTTCGT CATAGCAATAGAGTAGCCGCGGCGCCTAAGGCTACGAAATAAGCTCATTGAACTGTAATG GAGTGTGCCTAGTTGTTTCTCTTCTTACCAAGACTCAAGAGGGATTTGGAGCTTCATCTA CGATTATTATACCTACCTACGTATAGACTATAATGGATAGTAAAAATAGAAAGTAATTTC TTTTATATCAACTTTTGTATTCATATGTAAGTACTTACTTATAATTATTTATTTTTTTAG GTGAAA
3'UTR (SEQ ID NO: 46) GTTAACAATAAACAATACTGTTAACCGTACGTAGTGTTAAATAATACAAATATATGTATT
TTACAATAAGATTTCCTGTTTTTAAATTATCCTCACGTAACGTATTGGGTATCTAAGCGG TTGAGCAGTGAGATAAGTCTACCCCAAAGTAGTCCGTTCATCAAACGACATAACCATCTC GTCATTTAGTTAAAACAAAAAAAGAATAATGATAAAAGATATAAAATTCTGTATAAGAAC CAATACCTAGCACCTTTCTCACATTGCCAAACATACATTAAAAAACAGTATTATGCTTTT TTCGCTCTTTTATTCTATATTTTATATTAAAAAATACCTATAGTAAATACTGTTTTCATC GTCGCCGGTATTCATACACGTAGTATACGTACCTAACTCTGAAACTACTGGTTTGAATTG AAAAA
[0236] > M. sexta- Serine Proteinase homolog 3 (MsSPH-3); AF413067.1
5'UTR (SEQ ID NO: 47) AGCGCTTTGGGAACGTTCCGTGGCACGATA
3'UTR (SEQ ID NO: 48) .0 GATGGTGGAATGTTTAAAAATACAAGAGTGTGAGGCAGTGATTAATGGTCTGTAATATCC
GATTCTTGCTGGCTTCTGTTTCGTAATTAACGTAAAATCTAATTATCATTAGAATATTTT GGAAAACCTTAGAGTTACCTTCGGAAAACTTCACCGTTTGCCGCTGGTGTGAAGATCTGT TCCCCCCATAAACTGATGGGAAATTGTATGTAGGTACTCGAGTTATAATTTCTTTTTATA CCTGCGTTTAAGTTCCATTACAACTTTCTAATTTTATGAATAAATATCTAATAAACGATT AACAAAATTAAAAAAAAAAAAAAAAAA
[0237] > M. sexta- Peptidoglycan recognition protein 2 (MsPGRP2); GQ293365.1
5'UTR (SEQ ID NO: 49) ACGGCATAAATTGTTAGGTCGTCTGAGAGGCGAGTGTTGTATTTTTAATTGCCAAAGAGG
CTGCGAAGTTCGCAATAATCATTAGAAAA
3'UTR (SEQ ID NO: 50) AATGACCAAAGATAATACGACTGTTTTATAATTTTTGTTAATAAATGTTGTTGCATTATG
GAAAAAAA
[0238] > M. sexta- Beta-1, 3-glucan-recognition protein 2 (MsPGRP2); AY135522.1
3'UTR (SEQ ID NO: 51) ATTAAAACTAACGAAAAGACTCGTTTCCATAGTGGTATATTTTCAACTCGTCAATTTCAG
GTACAAGCATGTTTGGTGGAAGGATATTATCGGCCCGAATAGGCACTGTACACCAAGTAC AACCATGACTCATCAAAGGTGTCGCGTTCTGGAATCACCTGTATTTCCAACAGGCCGGCA TAATTGTGTCGTCTAGCGAGGGATAACTCAATAGTCTATTTGAACGCCATTCTACTTACC ATTAGGTGTAGTGAAGATATTTGGGGCTAGACAAAATGGAAGGAAATTTATGAGTTGCAT CGAATGATTATCACTAAGTATTCGAATGATTCATTTCGTTGTTGGTGCAATAGGATGTCA AGGGTCAAATGGACATTAAAAGAGAGTTTAAGGTGTTTTTTTACAATTAACGTTGATCTA TCACTTTACTTGCCTATTAAGTAAAGTATTATTTTGAAAAATACAAAATACTTGAGTTTA TAGTATCTCTTGCGTTTATGGTACTGTCAAAAATATCAGTATTATTCATTTACAAAATTA AAATTTAATAATCTAAATTATTCTTCATGTAAATGATCATTTATACCTCTGCCTTGATTA TG
[0239] > M. sexta-Relish family protein 2A (MsREL2A); HM363513.1
5'UTR (SEQ ID NO: 52) AGAGTACGTTCGATGGCAGTCTGTCGAACATTGGTAGTTTCCCGTTTGAGTGTTGTTTAC
TCCCTTTGAGGGATTAGTTTATTCTCCACGAATATAAACATCGGAAAATCAAAAACTAAG TTGATAAAAAGTTGTGTGCTCCGGTATAATTTTTTGGTATCAGTGACGGACAAAGGTGAT ATAAAA
3'UTR (SEQ ID NO: 53) TATATTCATGAGAAGGGGACAACTAAGGATTGAATATCAGCAGAACTTGAGTTATGTTAA
TGAGGTATTTATATTAGCTTAATACTTAAAGGGGAAAATTCGATTAGCTTTTCAAATATA CTTAAATTTTAGTTTTTGTCAAATATCGGTGTTTCCCATTTTGATATTTTTTTATCCATA TTTTAATAATAAATATCTTGTCTTAGCAATTTTAATGGGTAATTATAAAGAAACTCACGC ACCATAGTTGCACTAAACTACAAAATTACACAA
[0240] >M.sexta-Dorsal (MsDor); HM363515.1
5'UTR (SEQ ID NO: 54) ATACAACGATATCACAGGCGTCCGGGGACGGACCAGTTTAAACAAACATTACAGTGAATA
GCGATGTGATTATTTTCTTGCTTGTATAGAGTTAAATTTTTAAATTAGATTTAAATATTA AAATATTTGCGAATAAAA
3'UTR (SEQ ID NO: 55) ACTTAACAATTACATCTATAAATCTCTCCTCTATCAACCTAGTGGGCGTCCTCTACAACT
ACCACGTCTGGTATACAAATAGTGACAACCTGAAATTGTGAGATTTAAATTGTGTAGTTA TGATAAATAAACAATAAATCCAA
[0241] >M. sexta-Toll receptor (MsTOLL); EF442782.1
5'UTR (SEQ ID NO: 56) GAAAAGTTATTCACAATATGACCCGAAGTGTCAGCGCCGCACATGCGCGGGAGCACTCGT
ACTTATACCAGACATTGTGACTAAATACCTAATTTTATGTTTTGCTCATCGCCCATTGCG CGACATTAAGCAGTATCCAGTAGTCAGTGATCAGTGTTACTACACTATTTTGCTTCTCAG CGATTAGTCAACACGCGCATATCGTTTACTTCAACAAACAATTATTACGCAAATCGTGAT ATTTGAATGTAGAGACACGAATCAAAGATTATTCGTTTAACGTTATTTTCGCGGGATGTT
.0 TCTCTGTTGGGGGGTACTGTTGCGTGGTGTGCAGTCAAAGGTTTATTTTATCGGGTATTG
TGCTCTTTAAGTGTTGACGCGGCGGATTATCGTCGGTTAGACTAATAAATCGTGTCGATT TATGTGTGACCCACTACGGTGTCGTGCGACGTG
3'UTR (SEQ ID NO: 57) ATCCAGAACTTGCGTGTACCCTTCGATATACAGGGCTATTTTGACATCGTGTTACTAAAT
GAAACCACATTCTCGTCTTCACCTTTGCTGACATTGTGCCAAAAATCATCCATTAATAAC GAATATTTCCACCAAAAAAAAAAAAAAAAA
[0242] >M. sexta-Scolexin A (MsSCA1); AF087004.1
3'UTR (SEQ ID NO: 58) TAGACCATACCGTTGTCATTTTGGGCTGTAGTGTATAGATAATAAATATAGACGCGTGTA
CTGGTGTGACGTACGGAAGTGGAGAGTTGGGAGCGACAGCTCACCGCTCACTCCACTCCC GGCCGCCGCGCGAGTAGCAGTGTCAGTGATTGCA AAAAAAAAAA
[0243] >M. sexta-Hemolymph proteinase 18 (MsHP18); AY672794.1
5'UTR (SEQ ID NO: 59) GCCAGACGTTTCAGTTTGTTTCGAACT
3'UTR (SEQ ID NO: 60) TTATGAATAAATACAATTTAATAAGCATTATTTATTTAA
[0244] > M. sexta-Chymotrypsinogen-like protein 1 (MsCTL1); AM419170.1
5'UTR (SEQ ID NO: 61) TACTTCGGGCTACACAATGTACGTGAAAGTA
3'UTR (SEQ ID NO: 62) TCTCCGAACCGAACCTGATCTTAACTGTAAATAAAATAAGCGATACCCAAAAAAAAAAAA AAAAAAAAAAAAAAAAA
[0245] > M. sexta-Dredd (MsDRD); Msex2.04297-RC; Currently annotated as Manduca sexta Caspase-6; HM234679.1 in NCBI
5'UTR (SEQ ID NO: 63) TTGGTAACCCTGTGGGAAGTCCCAAAGTCGAAGCTCGAGAACAGTTTTAAGCAGAGGTT
TTAAGTCTAATTTATTGGTCTCTGGTTTTAAGTTAGTGTCATGTGTCAACAATAATAAA GCTTTATTTTTGGCTCTATTTATAAGTAAGGTATCATCCTTTCAGCGTTAAAACAAAGG GCGTAGCTCATGTTCATGTAGTTCTTTTAAATAAGTGTGAACTAAGTGTATATGTACAT TTATTACTGGTGACA
3'UTR (SEQ ID NO: 64) CATAATATTTTTATTTAGATATATTGTAAGTACTAAGAATGAAGTGGTGATAGCTTAGTT
GGGAGTGGAATGGACAGCCAAGACAAATGTCAGCAAGTTCAAATCCAAAGGCATACACCT CTGACTTTTAAAAAAAAATATGTGTGTATTCTTTGTAAATTATCACTTGCTTTACGGTGA AGGAAAACATCATGAGGAAACTCTCAATAGTAAAGGTATCCCATTGCCCAGCAGTGGGAC AGTATATTATACAGGGTTGATATTATTATTATTATATACTATGCTTTGCTCATGCGCTCA TTCACTCCAGCATTAATGACCTTTCCTGGTATTGAAGTTCTGCTAAGTATTTTATGGGAT AAAAATACCCTGAATGTTAATCCACACACATTCAGTATGTCAAATTTCATCTAAATCCAT TTAGGTGTGTTCTATTTGATACTAAGTTCCAACAAAAATCTTCATAAACGTCAGGACCCC TCAGGAGCAACATTATTAATAAGATTAATAATAAATTTTCAATGCATAATAATTTTAATA TAATTTATATTACTTAATGTTGAAGTTGATATACTTAA
[0246] > M. sexta-Relish F (MsReF); HM363513.1
5'UTR (SEQ ID NO: 65) AGAGTACGTTCGATGGCAGTCTGTCGAACATTGGTAGTTTCCCGTTTGAGTGTTGTTTAC
TCCCTTTGAGGGATTAGTTTATTCTCCACGAATATAAACATCGGAAAATCAAAAACTAAG TTGATAAAAAGTTGTGTGCTCCGGTATAATTTTTTGGTATCAGTGACGGACAAAGGTGAT ATAAAA
o 3'UTR (SEQ ID NO: 66) TATATTCATGAGAAGGGGACAACTAAGGATTGAATATCAGCAGAACTTGAGTTATGTTAA
TGAGGTATTTATATTAGCTTAATACTTAAAGGGGAAAATTCGATTAGCTTTTCAAATATA CTTAAATTTTAGTTTTTGTCAAATATCGGTGTTTCCCATTTTGATATTTTTTTATCCATA TTTTAATAATAAATATCTTGTCTTAGCAATTTTAATGGGTAATTATAAAGAAACTCACGC ACCATAGTTGCACTAAACTACAAAATTACACAA
[0247] > M.sexta-Chitin synthase 2 (MSCHS2); AY821560.1
5'UTR (SEQ ID NO: 67) ATTACGCTTTGAGCAGCCACTTTGTAAACACTAGCCTATTAACTCCGTCACTGCTTCGGC
GATAACACTTCTGTATATCTGTTATTGTTTGTTAATTTTTGGCGCAAGGATATATTTAAT ATGTGCTAAGTTTGTGGCTAATTTATATTATGTTTCTACATTAAAGGTATGTATTAGTGA TTTTTTGTAACTTATTTAATTACAAACATTATAACATATTTTATTAGAAAAGGGTTTTGT ATCCTATGTAGCGCCTGTTTCACGATGCCTCTTGAAGTATACTGAAATGGGTACCCGTCG ATAGTGTTTGCTTTAATAGGCAACGCATAGTGAAAACTTTGTACTTTAACCTTTTCAAAA GCATGAATAATTAAAAAAAAATATTAGAATAATGTGAAAAAGGTGTACAGTGTGATAAAT ATGGAGTATACAATATTCTTGTTCCAGTTATAAGATAAGGCACA
3'UTR (SEQ ID NO: 68) TATTTGGCCAAAATTGTGAACTGTAAACATGCAATAAAATGATGAACG
[0248] > M.sexta-Beta-fructofuranosidase 1 (MsSucl); GQ293363.1
5'UTR (SEQ ID NO: 69) TGAGAACAAAAACATTAGCTCCGCGTTTAAAA
3'UTR (SEQ ID NO: 70) TTATCGTGCTAAAGTAATAGTTATTGTTTCACATTATGTTCAAATAAAAAAGTAATTATT
TATGTTCAAATAAAAAAGTAATTATTTAAGTGTCTTGTAAATCTGCGAATAAATATACTT AATGTATAAAAAAAAAAAAA
[0249] > M. sexta-Sickie (MsSck); Msex2.03324RA (SEQ ID NO: 71) ATGACGACGTTGACTGCAAATAGGCAAGTGTCGGTGTTGCAGACGTCGATCCCGCTGCCG
.0 GCGTCGGCGGCGGCGGTGCGGCGCGCTCCGCCCGACAAACGACCGCTGCCCGCCACACCA
GTACATGCAGGTGGCAAGAGTGGTCTTTCTGCGAGTTCCAGCCGCTCCACCAGTCCATTA GGACAGGGTGCAGTCAGTTTTATACCACGTGCGCCTAATTCATCATCTGGATCCCGGCCG AACTCGTCCCTCCTTGCGCCCAGTAGCAAAATACCCTCCACCGCAACACAAGGGCAACAG CATTCACAAACACCCACAGCCCAAAATGGAACACCCACAAAGCAGTCTATGCTTGACAAG TTAAAGCTATTCAATAAAGACAAAATTAGCAATGAAAAACAAAACAGCAAAAGCACTGCA GTATCAAAACGCACCAGTTCTTCCAGTGGTTTCTCATCGGCAAAGAGTGAGAGATCAGAC TCGAGTTTAAGCCTTAATGAGTCTTCTAACATACCCAACACACATATCAAATCGTCTAAT TTAATAGGACCTAAAACAATACGGCAACAAAGCGATACTTTATCAAAAGACAAATCTGGT AAAAATTTAAAACCTAAGTTAGTTAATTCTAAATCTCCTAGAGATTCTACAACTAACTTG AATAGGGCTAGTAGCAAAACAACTAATAATGATAAATCGAGGAATAGTCCAAAACTTCCT GCTAGGGATAAGGAATCGAAACTAGCCACACCTAAAACCATGAGTAACACAAAACTAAAC CAAGTTGAAGATCAACCGAGGGGTTCCAAAACTAAAGTGGAATCGAAAATGGTAAAGCTT TCCGGTAGCCAAGTTAAGCTGAGTGAACAAAGATCCGATACTCCACAAGACTGTAAAGAA AATGTAACTCCTAATGCGAAAAATCATCAGAGTCACTCTCAAACACCATGTGGAGGACAG GGATTTGGAACTCCAAACCATACAGGAATACCTAAACCGACTGCCGCTGTCAAGGGAACT TTTAAAATATCAAAAGACGATAGACATGTTATACAGAAAAGTACAAATAGTCAATTAAGT CCTATACAAAGCAATTCTAGTTTAAATTCTACAAATAACATAAGTGCACTTCCATTCTGT AGAGAACAATCTAATTTAAGTAAGGATATAACTCAAAAGCAGACACTAGCCGTATCCCCG ATGCCGGTTATGAGCACAGGGAATCAAAATCATAGTTCGCAAATGTCTGAAAGTTCACAC TCCAATTCTACGCACAGTACTACAGGCCAACATTCGAACTCTAGCGATAGCAGCGTTATA TACCGTCCTTCTAGCGAATCAGGTTCTGAAATGTCGAAGACTGCTAGTCACAGTGTGGCA TCAAATAAACGGCTAGACATGAATACTACTTATATAAATGACGTTATAAACGAAGCGGAG ATATCAGAAAAGGAGGCAGCACAAAGACGTCACGTCGATAATTCAATCCCCAAGCAATCG TTTGATCCCAATAAAACGTTAACCGAAATAAGTAGGAGGCTCGAAAGTGATAGTAGATCT AGCACACCGTCGCATTGTCGTGATAATTCTTTAGGCGAAGACGAGAATCCAATGATGAAT GTTTTACCAATGAGACCGTTACTTCGCGGATACAACAGTCATTTAACCTTGCCCATGAGA ACTTCAGGTTTGGCACAGAAAAATATAAGCGTTTATCCACACCACGCAAATACTGTAAAA GCAAACTTTGGACGGGAAAACATAGGTCTTCGCGATCGTATAAACTATGGGCCTGGTTTT TCGAATCCTGATTATTGTGATCTGGAAATTGCTTCGGGATACATGTCTGATGGTGATTGT TTGAGAAGAATTAATATCGGTGAGATGGAATGCGGGAGGAACAACGACATGATGGACGGC TATATGTCCGAGGGTGGTGCTTCACTCTACGGTCGTCGGATGAACTACCAGCAACCACAA T TTCAACAAATGGACGAAAGACGAAGTGGTGGTCGCAATAGAGGCATGGAGGGTGGTAGT GGTGTAGTCTACAGAGTAGTGGGTCGCACACGCAGCAAAGCCGACTCCGGACAGCAGACC GAGCGCCAGCCTCAGCCTTCCCGGCAGGACACCACGTGGAAGAAGTACACTGACTCGCCC GGGAATCAGCCAGCACCCGCGCCTCCTAGTCCATCTCACTCGCGCAAGGGTGAGAGACGA ACTGGCCATCATTCACCGCAGCACCACAAACGAGAAAAACTCACTGCAGCTCAGCAGCTT GGGATCGCACCTCATCCTCAGTATGCCGCTCCAGCTCAAGCAGCTAACCCTGCTGGTCAG CATTCCTCTAGAAACCCAGGCCCACAGCTCCAGTCTCCGAGTGGAGGGTCGCGTCCCTCC AGCGTCTCTAGCGGTGGTAATGGCAGTGCCTGCTCGTCGAAGGCTAAAGTTCCTCAAAAT TTTGGATATGTGAAAAGACAAAACGGTGTACAGCAGCCGGCACCTCAGGCCAACGGTCCA CCGCCGCAACATGGAGGCCACACTGGAAGAACCGCCCAGGTGTCTGCAGTGCCAAGAACT AAAGTCAAAGTTTCGGGAGGAACCCAGACATGCACACAGGATTTACAGATTCACAAAAAT GGTATCGGACCTAAATCGTATTCTCTGGGCGGCACCGCGGCTGCCCAACTGTCAGCGTCA GTGCGAGAGAGACTGCTCGGGTCACAATCACTACCAAAACCTGGAACACACGAATTCGCA GCGTTATTCCATCACCACAGAATTGCACCCAGAGGCGGCATGAAGATCAGCGATGGCAGC CTTTCTGACACACAGACATACTCTGAAGTGAAATCAGACTATGGGATACCCTACGCGCCC TGGCTCAGGCATAGCAACACATACTCGGCGAGCGGGCGGTTGTCAGAGGGAGAGTCGATG GAGTCGCTCACGTCGCTGCACTCGGCGCAGGCGCAGGCGCACAACCACACCTCCTCCCCT AACTCCCACCGCAGCTCACTCACACACAATAAGCTCATCATGCACCGAGATGCACAGGGC TCCAGGCTTAACAGGAGCAACAGCATTAGATCGACTAAGTCAGAAAAATTATATCCGTCG ATGCTTCAAAGGTCTTCAGAGAGCGATTATGAACCGTATTATTGTTTACCTGTGCAGTAT GGACCAACAGGTCAAGGTCTAAACTATGGTGTGTCGGAGCCGCCGTCGCCGTCGCCGCGC TCGGCGCTGAGCCCGACGCACGCGCCCGGCAACGTCATGCACACGCCGCGCCACTCACAC CACTACCCGAAAAAGAACGACGATGTGCACGGTTCGACGGCGTCGCTAGTGTCGACCGCC TCCTCGCTGGCTGCCGGCGCAGGCGCAGAAGAGAGGCACGCCCATGAGGTGCGAAAGCTG AGAAGAGAACTGGCCGATGCGAAAGAAAAAGTTCACACTTTAACAACGCAGTTAACAACC AACGCGCACGTGGTGTCCGCATTCGAGCAGAGTCTGTCGAATATGACGCAGCGCCTGCAG CAGCTCACCGCAACCGCCGAGAGAAAGGATTCGGAACTGACGGAGCTTCGTCAAACGATT GAGTTGCTGCGGAAACAGTCGATTCAAGCTGGTCTGACTACTGCTCACATGCAGTCCATG GGAATCCGTGCCGATGGCGTCAACGTCACTGGACAGCAGGACCCAACAACCCAAACGCAG CAGTCATCACCACAACGTAACGCACAAACGGGCAACGGTGCGATCACGCGGCATCTATCG ACCGACAGCGTGTCGAGCATCAACAGCCTGAGCAGCGGCTCCTCAGCGCCTCACGACAAA AAACACAAGAAAAAAGGATGGCTACGCTCATCGTTCACGAAGGCATTCTCACGGAACGCG AAGATATCTAAAACAGCGAAGCACTCGTCCCTCGGGCAGCTGTCGTCACAGGACAGCTCG
.0 TCGGGCTCGCATCACTACGACGACCCGCACACGATACGAGAGGGCAGTAATGAGAACAGT
CTAGAACATTCCCACGAGGCTCTTCCTGATACTAAGGAGAAGCCGGCGCCTGTCAAGCCT GAAGAACAAACCAAAGATAACAAAGAGGAATCTGTCTTGGTGGATGAGCTGAAGCGGCAA CTGCGCGAGAAGGATCTCGTGTTGACCGACATCAGACTGGAGGCGCTCAGCTCCGCGCAT CAGTTGGAGAGTCTCAAAGACACCGTTATAAAAATGAGAAACGAGATGTTGAACCTGAAG CAAAACAACGAGCGTTTGCAGCGGCTGGTGACGTCACGCTCGCTCGCCGGCAGCCAGAGC TCGCTCGGCACCGGCGGCTCCGCCGTCGAGGACCCGAGGCGGTTCAGCCTCGCGGACCAG GCCACCATGCACCAGGCGGCGATCGACATTCACGCCCAGCCTCTCGACCTCGACTTCAAC TGCATGTCAGCCACACCAACCATCGACTTCTCGAAGAAAGGTTCACCCAAGTCCGGCATG GTGGAGCCCATATATGGGAATAAGGCTGCCTGCGAACTGAACGAGAACAATGAGACGCTG CTAGGAGCGTTGAATGGGGCCAGCGATTTGTTCTCCAACGGACTCAATGCTGGAGAGCGG CTGAGTGGAGATTACGATATAAACAGCGTGTTACCGCCGCCTAAAACACGCGAGCTTGCT ATTGGTGAAAGCTACTCTGATATTGGTGTAGCAGACAGCCAGGGAGACACGACTGATGGA AAGAAAATAGCGATAGCTGTATATTTAGGTCAACCCGAAACATTCCAAAGATATTTCGAA GAGGTCCAAGACACATTGACGGAGTCCGAGTGCAGATTCTACGCGAAGCAATCGGCAAGC GCGTACAATCATTTCGAGAAACAAGCCAGTTTCGAATCGCCGAGAATGTCCACGAATCAC AGTCCAGAAGTGGAGACACAAGACTACCCGCAAATAAATAAGTCGAACACGAATAGCCTT AAAAGCAACAAATCTACGCACAGTAGCTCGTATAAGAATGTTTATAATAGTGATTCGACA ATAAACTGCAACGAGTACACTATAGCGTACACTTATATATCTGGCAAGACGACTTGGCAG AATTTAGATTATATAGTTAGGAAGTCCTTTAAGGACTACTTGTCTAGGATAGATTTGGGC ACGAATCTCGGTCTGAACACTGATTCTATAACGTCGTACCATTTGGGTGAAGCGACGCGT GGGCCGGAGATCGGCTTCCCGGAGCTGTTGCCATGCGGGTACATCATAGGGACCGTGAAC ACGCTGTACATCTGCTTGCAAGGAGTCGGCAGTCTGGCGTTCGATAGCCTTATACCAAAA AATATTGTATATAGATACGTTTCTCTGCTATCGGAACACAGGCGAGTGATACTCTGTGGT CCCAGCGGCACCGGCAAGTCATACTTAGCGGCGAAACTCGCTGAATTTTACGTCCAGAAA ACACAAAGGCGCGGCAATCCCGCCGAAGCTGTAGCTACATTCAACGTGGACAGAAAGTCG TGCAACGAGCTGCGCGCGTACCTGGCGAACATCGCGGAGCAGTGCGGCGCGGCGGCCGCG GGCGAGGAGGCGCCGCTGCCGTCCGTCGTGGTGCTCGACAACCTGCAGCACGCCTCCGCG CTCGGCGACGCCTTCGCGGGGCTGCTGCCGCCCGACAACAGGAACATGCCCGTTATTATC GGTACCATGTCCCAAGCGACGTGCAACACCACGAATCTCCAACTACATCACAACTTCAGA TGGCTCCTCACCGCTAACCACATGGAACCCGTCAAAGGATTCTTAGCTAGGTATCTCCGA AGAAAGTTATTCTCGCTGGAGCTGCGGCTGGGTCGGCGCGAGCCGGCGCTGGCGGCGGTG CTGGAGTGGCTGCCGGGCGTGTGGGCCGCGCTCAACGCCTTCCTCGAGGCGCACTCCTCC
.0 AGCGACGTCACCGTCGGGCCGCGGCTCTTCCTCGCCTGCCCCATGGACTTGGAGGCCAGC
CAGGCATGGTTCGCAGACGTGTGGAACTACAGCATAGTGCCGTACGCTTCGGAGGCGGTG CGCGAGGGCATCGCGCTGTACGGCAGGCGACGACACGCCGCCGTCGACCCGCTGCAGCAC ATCAAGTCTACATACCCCTGGAGAGAACCCAATCACTCGCATACTTTGAGACCCATAACA GTTGATGATGTTGGCATCGAAGAGTCAAGCCAAGACTCCGCCGTGAACAACAATCAAGAT CCTCTGTTGAACATGCTGATGCGGCTACAAGAAGCGGCGAACTACAGCGGAAACCAAAGC CAGGACTCTGACAACGCCAGCATGGACTCGAACCTCACACACGACAGCTCTGTAGGGAAC GAGCTTTAA
[0250] > M. sexta-Akirin (MsAki); Msex2.12479-RA (SEQ ID NO: 72) ATGGCGTGTGCTACACTTAAAAGAAATTTGGATTGGGAATCCATGGCGCAATTGCCTGCT
AAAAGGCGAAGATGTTCGCCATTTGCTGCAAGTTCTAGCACAAGTCCTGGATTTAAAGTG TCTGAAACCAAGCCATCTACATTCGGAGAGGCCGTTAGTGCACCTGTGAAAATGACCCCA GAGCGCATGGCTCAAGAGATCTGCGACGAGATCAAGCGGCTGCAGCGGCGCCGGCAACTG CGGCTGGCTGGCAGCTCCGCCGCTTCGTGCTCATCGTCGAGCGGCAGCGAGGGCGACTGC TCGCCGCCACATCGCTCCTCGCACACTTCGCACAAGATGCACAACCGTGCGCTCTTCACT TTCAAACAGGTGCGCATGATCTGCGAGCGGATGCTGCGCGAGCAGGAGGTGGCTCTGCGC GCGGAGTACGAGTCGGCGCTCAGCACCAAGCTCGCCGAGCAGTACGAGGCGTTCGTGCGG TTCAACCTTGATCAGGTGCAGCGCAGACCCCCGCCCAGCACGTGCATGCCCCTCGGCATG GACGCCGAGCATCACATGCACCAGGACCTCGTACCTAGCTATCTGTCCTAA
[0251] > M. sexta-Cactus (MsCac); Msex2.02793-RA (SEQ ID NO: 73) ATGAGTGCCAAAAAAGGATATGAAACGAAGATTGTCGAGGAAGAAAACATGGATTCCGGA
ATTGTGTCTGGTGAATTGGAATCTTATGAGATTTCGGGTGAAGTGGATTCGGGCGTGATT GATTGTGATAAGAAATACGAAGGGGTTCCAAGTGAGGTGTTGGAATTGACGGACAAGTTC AAAAGTGTAAATGTGAGAGAGAAGAGCTGTCCTGATGTTCCACCACTGGCGGACCTGTTC CACCCTGACAACGACGGAGATACACAACTACACATTGCATCGGTACACGGCTGCGAGAAA TCAGTGAGCACGATCATCAGGGTGTGCCCTGACAAGGAGTGGCTGGACCTGCCCAACGAC TACGGCCACACGCCCCTCCACCTCGCGGTGATGAGCGGCAATGCCGTGGTGACAAGGATG CTGGTGATAGCCGGCGCTTCGCTCGCTATTCGCGACTTCATGGGAGAGACGCCCTTACAC AAGGCGACCGCAGCGCGAAACCAGGAGTGTCTCAAAGCCCTGCTTGCCCCTGTACCGGAA CAGCCCAATAGGAAATTGTCTTCAATACTCGACCAGAGGAACTATAACGGTCAATGTTGT GTCCACCTGGCGGCGTCAATTGGAAGCGTAGAGACGCTACAGACCCTGGTCTACTACGGA
.0 GCCGATATCAATGCCAGGGAGAACCTGGCGGGCTGGACGGCGCTGCACATCGCGGCGCGG
CGCGGCGACGTGCGCGTGGTGCAGTTCCTGCGGTCGCGCTGCGCCGGCGCGGCGACGCGG CCGCGGGACTACGCCGGCCGCACGCCGCGCCGCCTCGCGCGCCGCACCAAGGCCGCCGCC GCCTTCGACGACAAGGACGACAGCGACTCCGACTCCGACTCGGACGATGATGATATGTAC GACAGTGATAGCGAGACGTTGTTCGAAAAACTCCGCGAGAGCCTGAGCACGTCGATCAAC GTCGCCTGA
[0252] > M. sexta-Gloverin (MsGLV); G1110649240 (SEQ ID NO: 74) CCACGACAACCACTGATGAAGTTATTTTTTATAGCAATTCTTTTCGCTGCCATCGTCGCT
TGCGCGTGCGCTCAAGTGTCGATGCCCCCGCAATACGCTCAGATATATCCAGAATATTAC AAGTACTCCAAACAAGTCCGCCATCCCAGAGACGTGACCTGGGACAAGCAAGTCGGCAAC AATGGGAAGGTCTTCGGAACTCTGGGACAGAATGACCAGGGTCTTTTCGGTAAAGGAGGC TATCAACACCAATTCTTCGATGATCACCGCGGCAAACTGACAGGACAGGGTTACGGGTCC AGGGTCCTCGGACCTTACGGAGACAGCACCAACTTCGGCGGCCGGCTTGACTGGGCCAAC AAGAATGCTAACGCTGCTCTTGATGTGACCAAGAGCATTGGCGGTAGGACTGGGCTGACT GCCAGTGGATCAGGCGTGTGGCAACTTGGGAAGAACACGGATTTATCTGCGGGAGGCACT CTGTCTCAGACGCTTGGACATGGGAAGCCTGATGTCGGCTTCCAAGGTCTCTTCCAGCAT AGATGGTGA
[0253] > M. sexta- Beta-I tubulin (MspTub); AF030547 (SEQ ID NO: 75) ATGAGGGAAATCGTGCACATCCAGGCTGGCCAATGCGGCAACCAGATCGGAGCTAAGTTC
TGGGAGATCATCTCTGACGAGCATGGCATCGACCCCACCGGCGCTTACCATGGCGACTCG GACCTGCAGCTGGAGCGCATCAACGTGTACTACAATGAGGCCTCCGGCGGCAAGTACGTG CCGCGCGCCATCCTCGTGGACCTCGAGCCCGGCACCATGGACTCTGTCCGCTCCGGACCT TTCGGACAGATCTTCCGCCCGGACAACTTCGTCTTCGGACAGTCCGGCGCCGGTAACAAC TGGGCCAAGGGACACTACACAGAGGGCGCCGAGCTTGTCGACTCGGTCTTAGACGTCGTA CGTAAGGAAGCAGAATCATGCGACTGCCTCCAGGGATTCCAACTCACACACTCGCTCGGC GGCGGTACCGGTTCCGGAATGGGCACCCTCCTTATCTCCAAAATCAGGGAAGAATACCCC GACAGAATTATGAACACATATTCAGTTGTACCATCACCCAAAGTGTCTGATACAGTAGTA GAACCTTACAATGCAACACTGTCAGTCCACCAACTCGTAGAAAACACCGACGAAACCTAC TGTATCGACAATGAGGCTCTCTATGACATCTGCTTCCGCACGCTCAAACTTTCCACACCC ACATATGGCGACCTTAACCACCTGGTGTCGCTCACAATGTCCGGCGTGACCACCTGCCTC AGGTTCCCCGGTCAGCTGAATGCGGATCTCCGCAAGCTGGCGGTGAACATGGTGCCCTTC
.0 CCGCGTCTGCACTTCTTCATGCCGGGCTTCGCTCCGCTCACGTCGCGCGGCAGCCAGCAG
TACCGCGCCCTCACCGTGCCCGAACTCACCCAGCAGATGTTCGACGCTAAGAACATGATG GCGGCGTGCGACCCGCGTCACGGCCGCTACCTCACCGTCGCCGCCATCTTCCGTGGTCGC ATGTCCATGAAGGAGGTCGACGAGCAGATGCTCAACATCCAGAACAAGAACTCGTCGTAC TTCGTTGAATGGATCCCCAACAACGTGAAGACCGCCGTGTGCGACATCCCGCCCCGTGGT CTCAAGATGTCGGCCACTTTCATCGGCAACTCCACCGCTATCCAGGAGCTGTTCAAGCGC ATCTCTGAACAGTTCACCGCTATGTTCAGGCGCAAGGCTTTCTTGCATTGGTACACCGGC GAGGGCATGGACGAGATGGAGTTCACCGAGGCCGAGAGCAACATGAACGACCTGGTGTCC GAGTACCAACAGTACCAGGAGGCCACCGCCGACGAGGACGCCGAGTTCGACGAGGAGCAA GAGCAGGAGATCGAGGACAACTAG
[0254] > P. xylostella- Peptidoglycan recognition protein 2 (PxPGRP2); ACB32179.1 (SEQ ID NO: 76) ATGACGTTGTCTTTTGGCGTGTTTCTGCTGATATCTTCAGTGTTTTGTTGTTGTGCTCAT
GCAGGGTGTGGCGTGGTGACCAGACAGCAGTGGGATGGGCTGGACCCGATACAGTTGGAG TACCTGCCCCGGCCCCTGGGGCTGGTGGTGGTCCAGCACACCGCCACCCCCGCGTGTGAC ACTGACGCCGCGTGTGTGGAGCTGGTGCAGAACATACAGACCAATCATATGGATGTGCTG AAGTTTTGGGATATTGGACCGAACTTCCTGATTGGTGGGAACGGCAAGGTGTACGAGGGC CCTGGTTGGCTGCACGTCGGCGCCCACACTTACGGCTACAACAGGAAGTCTATCGGGATC TCTTTCATTAGGAATTTTAATGCTAAGACCCCAACAAAAGCAGCGTTGAATGCGGCTGAA GCATTGCTGAAGTGTGGAGTGAGAGAAGGACACCTGTCTCACTCATACGCAGTGGTCGGC CATAGACAACTGATCGCAACAGAGAGCCCAGGCAGGAAACTGTACCAAATCATCAGGCGC TGGCCAAACTACCTCGAGGATATTGATAAGATTAAAAACAACAAGTAG
[0255] > P. xylostella- Immune Deficiency Protein (PxIMD); Px003008 (SEQ ID NO: 77) ATGTCTATCCTAAAATCAAAGTTATTCGAAACTATTGCAAAAAGTTTCAAGTCTGATGCA
GTCCCGAAGCCACCTAGAGAACCGGTAGAGACTACAGAGACATCACAAAATAATACCGAA AATCAACCTTACAATGTCGAAGAAGAGGAAATACCCGAACCAGAAAAGCCTAAGAAAGAA AAAAAGAATCCCAAGCCTACCAAAAAAACTTTCTTTAATCGTGACAAAACTAACAAACAC
.0 GACGATACCCGCAAACATACAAAATCCGGAAAGGACCAGACATCAATTAATACTCAAGGT
AACTTGAAAATTATACTTCCTGTAACTTAAACGGCCCGTTGACCCCAGTTTACCTTTCGC CTTTCTTGATATATTTTTGTAATCCAGCCTTACTTTGGTAATACATACTTGCCCCACTTG TATTTAGTTAATGGTGGCACTAGCTAGATAGTAATGTTAAATGATGATAAGCAGTAGTGA TTCATCATTCAAATGTATCATTGTCCTTTAATGTTAAGCGCAAATAGATTTTCATTGTTC TCCCATGTGCTTCATGTTTTATGTATTTATAGGTAGGTACTTAATGTTTTATAAATATTT TTTTGTTAATTGGGAATCCCCAGTCCCCATTGTCTGGACCAGTTTATATATAATTGAACT AACAAGAGTGTGCTTTAAATACTATTCTCTGCAATTATGATAATTAAACAACATGAATTT CTCTTCACTTCCCTTCTCTTATTTAAATAATATTGTAGGAAACTGTAATAACTAATACAA GATTATAAATTTCATTCTAGCAACTGGTGATGTAATCCATGTGGTAAATTCCAAAGATGT GCAGGTCGGCCATCAGTATGTGTACAACATGGGAACTCCCGGAGCTAACTCACAGAAGAA TAACCCATTTGATGATGAAGAAACAGTAGAAAAGACAAATCTAATAACTCTGGTCATGGA AGCAAAAATTATGGTAATAACACATTTTTAACTAGGCATAAGGTCATAATTTAGCCAGAA TCATCAGCTTGTCCTGTGGCTCTTGTTGAGCTGGTGGAAAGAATACATAGGTAATGAATA TTTTGATCAATACTCATTGCAAAAATCACAATAATGCCATTGAAAATCTATAACATGTTC CTTAAGTATCACTTATCATCAATCCAATTAAGTCATCACACAGATCAATCGGTTAGTTCT GTTTATTTACTTCTTTCAGCTGGAACATGAATACATGGACTATGTCTCGAAGAACCTCGG CAGGAACTGGCACAGCTTCTTCAGAACGCTCGGCTTCACGCGGGGGCGCATCGAGACTGT GGAACTGGATGAGGGCAGAAATGGTGTTGCAGAGGTATGAAAAAAACATACATGTTAATT TGTGTTGTTTGGCTGATGTGGCAACTAAGTTCGTAACTGCTATGATCAACTGTTTGTGTC ATAGGTATTTTTTTTGCCCTTCCACACATCTGAGGTAACAAGGACACCTCCTGCACTCAA AACACAACTGCTATGTCCTACTGATGAGTCCTAGGAAACTCAGAAACCATCTGTTTCCCC CATTCTCAATTCCATAAAGCTAAAAAGTGGTAGTTACACAATTCACAATTCATAAACATG CTTTGTCATAGTTAGAAAAGCAGCTCAGCTATGAGGCATCCACTCCACAGTCCACTCACG AAAGCCCTCTTTAAAAACATAAAATCATCATCATCAGCCCTCAATTGCCCACTGTTGCAT ATAGGCCTTCTTTTGATTATGCCAAGTTTTTCGTTTCTCTAGTCCACAGACTTGACATAG TTGTCACAAATTTATTCTACGTACTTGTATTTCCAGGTGCGCTACAAGTTGCTCCTGGAG TGGGCCCGCACCGACGAGGACCCCACGCTGGGGCGGCTCGCCACGCGGCTGTGGGACGAG GGAGAGCGGCAGACCGTCAAGGAACTCGCCATCTTGTATAATAATAATTTCAAGCAACAA TGTTGA
[0256] > P. xylostella- Relish (PxRel); Px002858 (SEQ ID NO:78) ATGGTAGTGGTGAAAAGCCAATTAAAAATGCAAAGTGACCAGGACACGGACTCGTCCACT
.0 GCCGGTGCTTCGCCGCGCAGCTTCTACATCGAGTCGCCGCACAGCTCGCCGGGACAACAA
GTGCCTTATTTAACTAATTACATGACAGTACTGTCCTGTGCAGATAATAACTTAATGGAT ACAGGAAGCAATGGACCATTCCTGAGCATCACGGAGCAGCCATGTGACCACTTCAGGTTC CGCTACAAGAGCGAGATGGTCGGCACCCACGGCTGCATCGTCGGCAAGACCAGCGCCAGC AACCGCACCAAGACATACCCTTCTGTTGTTCTGCTCAACTACAAAGGCCGCGCCACCATC AAGTGCAGCCTCGCGCAGCACAACAACCGCAAGCAGCACCCGCACCAGCTGGTCGAGGAT GACCAGGAGCGCGACCTGAGCGCCGAGGTCAACCCCGAGAAGGGCTATGAAGTTGGATTT CGTGGCATGGGCATAATACATACAGCGAAGAAAGATGTTCCGGCACTCCTATACAAGAAA TTGAGTGAGAGACTGCCACATTTCAATGCCCGTGAGCTGAAGGCCCAGTGTGAGAACGAG GCGCGCAGTATAAACCTCAACATCGTGCGCCTCAAGTTCAGTGCGCACAATGTCGACACG GACGAGGAGATATGCGCTCCGGTGTTCTCGGAACCTATCCACAACATGAAAAGCGCCGCG ACGAACGACCTGAAGATCTGCCGCATGAGCCGCACGTCGGGGCGCCCGCGCGGCGGCGAC GACGTCTACCTACTCACCGAGAAGGTTAACAAAAAGAACATCGACATTCGCTTCGTGCAA CTGGAGCGCGGCGAGGTGTGCTGGACCGGCAAGGCCAGGTTCCTCATGAGCGACGTGCAC CACCAGTACGCTATTGTGATCAGAACACCAGCATACAAGAACCCCGAGATTACGTCTGAC GTAAAAGTGTACGTAGAACTATTCCGCCCGTCCGATGGCCGCTCCAGCGAACGCATAGAG TTCACGTACAAGGCAGAAGAAGTCTACAAGCAAAGCAAGAAACGGAAGGCCAACTCTTAC TCCTCTATCGGAAGTTCATCTAGCGGTAATTCTATCAAAAGCGTCAGTGATCTTCCAGCA ACTGTTATTATGGCCAATGAAATGAATGCGGCTAACAACAACTTTAGTAAAATCTCTTCA ATGCTGAGCCCAAACAATATACCAGAAATACCGACACAAACGACTGTGGGTCTATCAGAC GCTCTCTACGACATAACAGTGACAGAAGACCACCAAATGCACATCAGTCCCATGCTATGC CAACCAGTGGAAGAGTATCCCCTAAAGCTTAACTCGCAGGATATCATACAAGTGAATTTG AACTCTAAGGACATCGACCAACTGCTCAAAGTCAACAGTGTGCCTGATACCGATAAAGAC TTTGCTGACTTCAATTTTAGTGACTACTACAAGGCACTCGATAGCAACTTTTTAGCTGAT GGTGGTGGTGATAGCTTCAGTCAGTGTATCTTCAACTCTATGCAACTGAGGCCTGACTCT GGGAGAGGCACG
[0257] > P. xylostella- Toll receptor (PxToll2); Px006338 (SEQ ID NO: 79) ATGCCTAAAGTAATAATCGCCAGTTTTGCTTATATAGTAGGTGTGTTTGTCCTGTGTGCT
GGGCTAGAAACAAGTCCAACCTGTTCCAGCATCGAAGGACCTACTCCAGGTGAATTCATA ATACAAAGAGGTATCGTACCGGACAATGATTCAGCCACCACAGTAAGCTTCCGAGGCTGC CGAATATCTGACATTCAGCCGAGAGCATTCCATGGGCTACCTTCCCTGCAATACATAGAC TTATCAAGAAATAGCATTAAAAACCTGAAACTTGGCATTCTTAATGACGTCACTAGACTC
.0 ACTCATCTGAACTTGTCTTATAACTTCATCAGCGATTTAGAAGAGAGTTTGTTCAACCAA
TCGTCAAGGTTGGAGGTGTTGGATCTCCGGTGGAATAAGATTGAAGTTATGAAAGTGGGC GTTTTCAGTCCATTGAAGAGATTGAAGTATTTGGATCTGTCCGACAACGAAATAGTTGGG GCCAGCCTGAGCCCCGCTATGTTTGATTCCTGTAAAGCTCTATCCACTATCAATTTTTCA AGAAATGATATGTCTGGTGCTCCCACTGATTTGCTTCGAGCTGTGGAGGTACTGGACACA CTGAAACTGGATGGGTGCTTTTTGAAACAAGTTCCGGAATTTGCTACGAGGAGCAATACC GGCACAATGAAGAAACTTATTTTATCATCGAACCAAGTGAGCACTGTGAAACTTACTACG TTCATCAGTTTAACAAACCTGGAGGAACTAGATTTGAGTTCAAATGTAATTTCAGAATTG CATGAAGACGTATTCAAGCCATTAAAAAACTTGAAAATTATTATTTTACGCTCGAACCGA CTGGAAAAAATTCCTGATAAGTTGTTTTATAATATGTTACGATTAAGGAAGGTAGATTTA TCTTTTAATTCTTTGATGATAATACCTGTGAATGCCTTTCGTTTTACGACGATAGAAATG TTGAATATATCGCATAATAAGTTCACATATTTAGTCGACAACTTTTGTTTGGAACTTAGA AACTCGGGAGTGAAACTGAAAAAGTTTTACTTCAACAGTAATCCCTGGCAGTGTCCTTGC TTGAGGGATTTATTAAAGGAAATGAAGACGTATAGAATATCGTACAATAATGCTAAATAC GATGGTAAAAATGCAGTTTGTATTTCAGGAGACATTATTAATACTTGCTTGAGACAACCT GATGTCAATGAACACTTCAATGATTTGTACTATTCTGACTAA
[0258] > P. xylostella- Cactus (PxCac); Px016665 (SEQ ID NO: 80) ATGAGTTTCAAGAAGGATTTCGACACCTCAAAGAAGATCCAGGAGGATGAAAACACAGAC
TCTGGGTTCCTATCTGGGCCGATAAGTGAGCAGCTGACCTCGGAAGATTGTGATTTAGCG GAGGAAAGTGAGCGTGCTCGCAGCAGGCTTAGTGAGGAAGATCCTGAGCCTGAGCTGCAG TTGGACAGTGGGCTGGACCTCTCGGAGTGTCTGTCGAGTGTTAAGCTTAGTGATAGTGCA GTGTACACACCCACCTCGCAGACCACCCCCACAGTCACTATAGGTGATGAGAAAACTCAT GACATCCCACCCCTCGCCATCCTGTTCCAGCAGGATGACGATGGAGACACACAACTACAC ATTGCAGCGGTACATGGGTGCGAAAAATCAGTAGGAACATTAGTAAGAGTTTGCCCTGAC AAAGATTGGCTAAATGTACCAAATGACTTTGGACAGACCGCCTTACACTTAGCAGCCATG AGTGGGCATGCAGTAGTCACACGCATGCTGGTGATGGCCGGTGCATCTCTTGGCATTCGA GACCTTGTTGGCAACACACCTTTACATGTGGCAGCCGCAGCGGGCTACGTCGGCTGTCTC CAAGCTTTACTGGCTCCTGCTCCAGAACAACAGCAGAGAAGGCTAGCATCCACGTTGAAC CAGAAAAATTACAATGGTCAAACGTGCGTCCATGTGGCTGCGATGGCCGGCCACGTCGAC GCGCTGCAGACATTGGTCTATTACGGAGCTAACATCAATGCTGCGGAGGGTCTATGCGGG TGGACACCTCTACACGTAGCGGCGGCGCGAGGCGACGTCGACACGGCTCGCTACTTGCTC GAGAAGTGCGCTGGCGTCGATCCCTCTGCCCTGGACTACGCCGGTCGTACGGCCAGGAAA
.0 C TGGCGTTGAAGAATAAAGCGGCCGCCCTGTTTGACGGCAGTGAGGGCAGCGAGGAGGAG
GATAGTGACAGTGAGGATGAGATGCTTCTGGAAAGCGACCAGAGTCTGTTCGACCGGATC CGTGACGGTATGAACGCCATCAACGTCGCCTGA
[0259] > P. xylostella- Dorsal (PxDor); PxOO0110 (SEQ ID NO: 81) ATGAACGCGCCCGCCGACTCCGCCGTGGTGACGTTCACCAACCTGGGCATCCAGTGCGTG
AAGCGGAGAGACATCGAGGACGCCCTGGCTGTGAGAGAGGAGATGCGAGTTGACCCCTTC AAGACCGGATTCAGCCACAAGAACTCCCCGCAAAGCATCGACCTGAACGCCGTCCGACTC TGCTTCCAAGTGTTCCTGCCGGACGAGCGATCCGGCAAGATCCGCCACGCGCTGCCGCCG GTCGTGTCCGATGTCATCTATGACAAGAAGGCCATGAGTGACCTGGTTATCACGAGGCTG AGTCATTGTTCTGCGCCCGCGCAGGGCGGCAAGCAAGTTATATTGCTGTGTGAGAAGGTG GCCCGCGAAGACATAACCGTAACCTTCTTCGAGAAGTCCGGCGAGCGCGTGACGTGGCAG GCGGACGCGGCGGACGTGTTCGTGCACAAGCAGGTGGCCATCTGCTTCACCACGCCGCCT TACCGCGACCCGCATGTGCAGGACCATGTGCAGGCGTACATCCAGCTGCGTCGTCCGACG GACAACGCGACGAGCGAGCCGCTCCCCTTCGAGCTGCTCCCGTCCAGCGCAGATCCGAAT TATCTGAAGCGAAAGCGACAGAAACCGATACAGAACTTCAGTCGGTACTTACAGCCGATC GATAGCGACATGAAGCAGCAGCTGCCGGACTATTTCCAGGACAACATGGCGCTGTCCAGC ATCCCCTCCGTGAAGCTGGAGCCCCGAGATAAGACTCCTCCTCACAACATGAGCAGCCCG CCGCTGCTGTTCCCCCCCGCGCACGCCGCGCCCGCACACCATGACCCCTACGCGTGGAAC ATGCAACTAGACAACATGCAGTCGGGTCTGACGGCGCCCGGGCCGAGCCGCCTGCCGCAG TACAGCCAGGACATGGCCTGGACCAACCAGATGGGCCACGTGTCCCCTATGCACCAGGCC ATGTCCCCTAACATGGGTCACGTCTCCCCCATGCATCAAGCCATGTCCCCAAATATGGGC CATGTCTCTCCCATGCACCAAGCTATGTCTCCAAATATGGTCCAGTCACCTATGGGTCAT GTGTCCCCTAACATGGGCCATGTATCCCCTAATATGGGTCATGTGTCCCCTAATTTGGGT CATGTGTCACCTAACCTGTGCCAGCAACCAATGGCTCCTATGGCGCAGCAGCTGATGGAC CCGTCCCCCAGCGACCCACCCTCCATCACGGGGCTGCTGATGGATCGCCCGGACCAGCCC TACTCCGGGGAGCTGTCTGGACTCTCCGCCCTGCTGGCTGAGGCAGCCCCCGCAGAGATG CTCAGCGATAGCCTCAACAGACTGTCTACGGGGGACTTGTTGAGACAAGTTGATATGTGA
[0260] > P. xylostella- Hemolin (PxHem); ACN69054.1 (SEQ ID NO: 82) ATGACTTTAATTTTCAAGAGTGTTTTATTTTTGGGCTTAATATTGACTACTTTTATTGTT
TCCGCTCAGCCTGTGAAACAAGATGGCGGCTCAGCACACGAAGAAATATTGTTCCGTGAG CACGGCCAGCCGGTGGTGTTGACCTGCGCGCGCGCAGACGACCCCAACCAAGGTGGCATT
.0 AGAACGTGGTTGAGAAACGGGACGCCATTAGAAGACGGCAAAATGTCTCCCGAAATAAAA
TTCCTCGACGACAAATCACTCTGGTGTTGCAGCCCTCACCAGCCGTGGAAGGGGTCTACC AATGCTTCACCGAAACCCATAAGGGCATTGCAACCTCCCCGAAATTCAGCGTGAAACAGA CTTATCTCAAAGCTCCAGAGACTACGCCTTCAGTCAATATCAAACCAGCAAAAGGCCTTC CCTTTAGCTTGGACTGTGACGTCCCTGAAGGATATCCGAAGCCTGAAGTGCAATGGTTCC TACAACACGGGAAAGATCACACCCTGATTGAGGCAATTATCAATAAACGGATCACACAGG CTCCGAACGGAGCTCTTTACTTCTCAAATGCTACAACCGAAGATGTGAATGTGGGAGACT TTAGATACGTCTGTATGGCGAGGAATGATGCGGTAGACTTACCAGTGGTGGTGTCGGAAG CTGTCATCACAGGTCTGAGCAGCGAGGGTGGTAAGGGTAGATTGGTGGAGCAGTACGTCA GTAAAGAAGTTAGGGCGGTTGCGGGGGAGACCACAGCGCTATTTTGCATTTTCGGTGGCA CCCCACTAGCCCACCCAGACTGGACGAAAGACGGCAAGAATGTGAACGGGGCGCCCGGCG ACCGAGTGACCCGACACAACAGGAGCTCAGGAAGACGACTCATCATCAAGAACACCACTC TAGAAGATGCTGGAACTTACCAGTGCGCCGTTGACAACGGCGTTGGGACTGAAATGCGTT CTGTCAAGGTTACTGTTGAAGCGAAACCTTCAATAACGATTGTGAATGAAGTAGCAGCGA AGCTTGGAGAAGAAGTCAAGATTTGCGAAGCGACCGGAGTCCCAACGCCAAAACTAACGA TAACTCACAACGCTAAACCGTTGGTTGCGTCAAATAACGTTGTTATAACCAACGATGGAG TTGTTATAAAGAATATTCAGGCTATTGATCGAGGGTATTATGGGTGTGATGCTGTCAACG AGCTAGGAAGCGAATTTCGTGAAACTTATCTAAGTATTGCCTGA
[0261] > P. xylostella- Sph-3 (PxSph3); XP_004922188.1 (SEQ ID NO: 83) ATGCAGCTTAATTTCTGTATCAATGTGATCGCGACCATATTGTTGATTGTGACTGGCGGG
GATTCCCAAAAACGAGTGGGTGACATTTGCATTGATCAATACACGAACACGTACGGAAGG TGTGTTTTCTCGGATCGATGTCCATCAGCTTTACGTAATTATCAACAGAACGGCATTCGG CCATCAATATGCACTTACAACTTCGACAATGCACTGGTGTGTTGTACTGAACGCGGAAAT ATTCTTCAAACGGCGAGGCCCCCGCCACCGCCTGATCAGGAAGACAGATTTCAGTCCTCT GCTGGAAACAACAACAACAACAATAAACCTAACATTAGAGTTAGCGAAAGAAAATGCCGC GAGTACAGCAAATCAGTAACGTTCACGGTGAGCTTCAGCTCGCTGCTGCCGGAGCCCGAG TTGCAGTCCATCTCGCGGCCGCGCTGCAGCCGGAGCGGCGTGGGGCTCGTGCTCGGCGGC CGGGACGCCGCGCCGGAGGAGTTCCCGCACATGGCAGCGATCGGCTTCGCATCAGCGGAA GGCTACGACTTCAAGTGCGGGGGGTCCCTCATCAGCGCGCGCTGGCCGCTGACCGCGCCC TGCGCGCGCGCCCGCGCCTCCAGCCGGCCCGTGGTGGCGCGCTTAGGAGATAGGAATATC AACCCGAAAGCGCAGGACGACGCCACGCCTGTCGACGTGCCAATCCGGAACATCATCGTG GACGTGGACCTCAGCAACAGCATCCGCCCCGCGTGCCTGTGGCCCGGCGGACCCTTCCAC
.0 GAGGATAAGGCTATAGCTACGGGCTGGGGGGTGGTGAACCAACGCACGCAAGAGAAAGCG
GACCTCCTCCAGAAGGTCTCGCTCACTCTGCTCGAGAACTCATACTGCGACCGTCTGCTG AGGAACAACCGCAACCGACACTGGCAGGGCTTCCGCGACTCGCAGTTGTGCGCCGGCGAG GTGCGCGGCGGCATGGACACGTGTCAGGGCGACTCCGGCGCACCGCTCCAGATCGTGTCC AAGGAGAACCAGTGCATCTACCACCTCATCGGCCTGACCTCCTTCGGCTACAAGTGCGCG GAGCAGAACAAGCCGTCGGTCTACACCAGGGTGTCGACTTACGTGGACTGGATAGAGTCT GTGGTGTGGCCGGAGGAGTATGCGGCTTGGGCGGCGGGGAGGAGTAAATAA
[0262] > P. xylostella- Transferrin (PxTrs); BAF36848.1 (SEQ ID NO: 84) ATGATAGTGAAAATAGCCATTTTGGTGATAGCAATAACGTTCAACGATGTGTCTGCGAAA
ACTTCGTACAAGATCTGCGTACCGTCTCAGTTCATGAAGGCATGTGAACAAATGCTTGAA GTGGAAACGAAGAGCAAAGCGATACTGGAATGTTTGCCGGCCAGAGATCGAGTGGAATGC CTGACCCTGGTGCAGCAACGGCAGGCGGACCTCGTCCCAGTGGACCCTGAAGACATGTAC GTGGCGAGTAAGCTGCCCAACCAGGACTTTGTGCTTTTCCAGGAGTTCCGGACCGATGAA GAGCCGGATGCGGAGTTCCGTTACGAGGCCGTCATAGTTGTTCACAAGGACCTTCCAGTT ACCAACTTGGACCAGCTTAAGGGCTTGAAGTCATGCCATACTGGAATCAATAGAAATGTG GGGTACAAGATACCACTAACGATGCTGATGAAGCGCTCCGTGTTCCCTGCGATGACAGAC CGCAGCATCTCTCCTAAAGAGAACGAGCTGAAGGCTCTCTCGACGTTCTTCAGCAAGTCC TGCATCGTCGGCCAGTGGTCGCCTGACCCGAAGACCAACACTTTCTGGAAGTCCCAATCC AGCAAGCTATGCTCCATGTGCGAGGACCCTGCCAAGTGCGACTACCCCGACAACTACAGC GGCTACGAGGGCGCGCTGCGCTGCCTGGCGCACAACGGCGGCGACGTGGCCTTCACTAAG GTCATCTATGTGCGGAAGTTCTTTGGGCTCCCAGTAGGCACAAGCCCGGCGACTCCTTCT TCTGAGAACCCGGACAACTTCGCGTACCTTTGCGCGGACGGGTCCAAGGTCCCTATCAGA GGAAAGGCATGTTCTTGGGCCGCAAGACCGTGGCAGGGGTTGTTGGGACATCAGGACGTT CTGGCCAAATTGTCGCCTTTGAGGGAGAAGATTAAGCAGCTGTCTAGAGCTGGAGCAGAA TCGAAGCCGGAGTGGTTCACCAACGTTCTAGGCCTCTCTGAAAAGATCCACTTGGTCGCC GACAACATTCCCATTCGTCCCGTCGACTATCTGCAGAAGGCCAACTACACTGAGGTCATC GAGAGAGGGCACGGCCCGCCTGAACCTGTTGTGAGACTCTGCGTGACGAGCTCGGTGGCG CTGGCGAAATGCCGCGCCATGTCCGTGTTCGCCTTCAGTAGAGACATCCGCCCCCGGCTG GACTGTGTGCAAGAGGCTTCGGAAAGCGATTGCTTGAAAAGTGTCCAAGACAATGGCTCA GACCTGGCGTCAGTAGACGACATGCGGGTAGCGTCAGCATCCAACAAGTACAACCTACAT CCAGTATTCCACGAGGTATATGGAGTCAGCAAGACCCCTAACTATGCGGTAGCTGTCGTC AAGAAGAATACTCAGTATGGAAAGATTGAGGATTTGAGGGGAAACGGTCCTGTCACAATC
.0 CTTTATGGAAGCTTCAGTGGCTTTGATGCCCCTCTGTACTACCTTATTAATAAGAAAATC
ATAGGCACTGAACAGTGCCTGAAAAAGCTTGGAGAATTTTTCGCAGCCGGATCTTGCTTA CCTGGAGTAGGCAAATTAGAGAACAACCCTACAGGAGATAATGTCGATAATCTGAAGAAA CAATGTTCTGGAGACAACAGCCCAATAAAATGCTTACAAGAAGACAAAGGAGACATAGCA TTTGTGTCAAGTGCTGACCTGAAAAACCTGGATGCCTCTCAATATGAGCTGCTCTGTCTA AACAGAGAGAACGGTGGGCGAGACTCAATAACCAACTACGCTACATGCAACATTGCCATG GCCCCATCCCGAACCTGGCTCTCAGCTAAAGACTTCCTGTCCGATGTGTCCATAGCACAC ACTCCGCTGAGCTTAGCACAACTACTGGATACCAGAAAGGATCTGTTTAACATTTACGGA GAGTTTTTGAAGAATAATAATGTTATTTTTAATAATGCTGCCACTGGACTGGCCACAACA GAAAAGATGGACTTTGAAAAGTTCAAGGCAATCCATGATGTTATCTCATCTTGTGGTGTC GCATAA
[0263] > P. xylostella- Transferrin (PxTrs); BAF36848.1 (SEQ ID NO: 85) ATGCTGCTTAGGACGATACATTTGTTGTTAATTGTTTGTTGTGCGTGGTGCTATGAAGTG
CCTCCGGCTAAATTGGAAGCTATTTATCCAGCTGGCTTGCGAGTGTCAATACCCGACGAT GGCTTCTCGCTATTCGCCTTCCACGGCAAGTTGAACGAGGAGATGGAGGGCCTGGAGGCG GGCCACTGGTCCAGAGACATCACCAGACCCAAGAACAACCGCTGGGTCTTCAGCGATAAA CAGGCCAGGCTCAAGATAGGGGACAAAGTGTTCTTCTGGACGTATGTTATCAAGAACGGA CTCGGGTACCGACAGGATGACGGGGTGTGGACTGTTGAAGGATTCGTCGACGTCGAAGGC AACCCGGTTGACCCCGCGAATGGACAACCCATCTCAGCGCCGACCAGACCTCCAACCCAA CCAGGCCGGGTGCCAAATGTCCCCATGCCGTGTGACATCTCAGTCACCACGGCATCAGTG CCAGGGTACATCTGCAAGGGACAGCTGCTCTTTGAAGACAACTTCAATGGGGCTCTGGAG AAAGGAAAGATATGGACGCCGGAGATTATGATGCCTGATGAACCGGATTACCCGTTCAAC ATCTACCTGAACGACAGGAACCTGCGCGTGAGGGACGGCCGGCTGTCTATCAAGCCCGTC ACGCTCGAGTCCAAGTACGGGGAGGAGTTCCTGGCCAAACTAGACTTGTCTGCCAGGTGT ACTGGTAACGTGGGTACTACCCAATGCAGCAGAGAGTCCATTGGGGCCCAGATCATACCT CCGATAATCACAGCCAAGGTTACCACCAAGAACAAGTTCAGCTTCAAGTATGGAAGGATT GAAGTGAGCGCCAGAATGCCGCGCGGTGATTGGTTGATTCCAGATATTCTGCTGGAGCCG AAAGAAAACCTTTACGGAGTACGCAATTACGCGTCAGGTCTACTCAGCATAGCCTCAGTC AGAGGAAACACTGCTTACTCGAAGACCCTCAAAGGAGGCCCCATACTGTGTGACAAGGAA CCGCAGAGAAGTGCCAAGTTGAGCGAAAAAGTTGGATATGACCATTGGAATAAAGCCTTC
.0 CATAACTACACCATGATTTGGGCACCAAGTGGCATCACCATGCTGGTGGACGGCGAGCAG
TACGGGGACATCCGTCCCGGCGACGGCTTCAGCCAGGACCCGGCGGTGAGCAGCGTGGTG GCCGCGCCGCAGTGGCTGAAGGGCACCAGCATGGCGCCCTTTGATGTTATGTTCTACATA TCCCTTGGTCTCCGCGTGGGCGGAGTGAACGACTTCCCCGACACTCCTGAGAAGCCGTGG AAGAACAAGGCCACTAAAGCCATGCTGAATTTCTGGAACGCCCGGGAACAGTGGCAGAGC AGCTGGTTTGAGGACACCACTGCACTCCTCATAGACTATGTCAGGGTTTATGCGCTGTGA
[0264] > P. xylostella- Gloverin (PxGlv); ACM69342.1 (SEQ ID NO: 86) ATGTACCGATTTGCAGTTATTTTATCTGTAGTCGCCGCGTGTGCCGTGGCTCAAGTTTCT
CTACCTCCTGGATATAATGATAAATACCCAGGCTTCTACAAATACTCCAAGCTAGCCCGG CATCCGCGACAAGTGACGTGGGACAAGAATGTCGGCCGTGGGAAGGTGTTCGGCACCCTC GGCGGCACTGACGATAGTCTCTATGGTAAGGCGGGCTACCGTCAGGACATCTTCAACGAC CACCGCGGCCACCTGCAGGGTGAGGCTTCTGGCACCAGGGTACTCAGTCCCTACGGAGAC AGCAGTCACCTGGGCGGTAGACTCGACTATAGCAACAAGCACGCCAACGCCAACCTGGAT GTCAGCAAGCGGATCGGAGGCGTCACTAGTTGGCAAGCAGAAGGCAAGGCTAGATGGCCG ATTGGCAAGAACAGTGAGCTATCAGCCGGCGGAATGATCAGACAAGACCACTTCGGCCAC GGGAGACCAGACTACGGAGTCGTCGGTGGGTTTAAATCTAGGTTTTAA
[0265] > P. xylostella- Chitin synthase 1 (PxCHS1); KX420688.1 (SEQ ID NO: 87) ATGGCGACGTCGGGGGGAGTGCGGGGGCGGCGGGAGGAGGGCAGCGACAACTCGGACGAC
GAGCTGACCCCGCTCCAGCAGGAGATCTACGGCGGCAGCCAACGCACAGTACAAGAAACA AAAGGATGGGATGTGTTCCGAGAGATCCCGCCGAAGCAGGACAGCGGGTCGATGGAGAGC CAGCGCTGCCTGGAGATCACCGTGCGCATCATGAAGATCCTGGCCTACCTGGTGACCTTC GTCGTGGTGCTGGGTTCAGGGGTGCTGGCCAAGGGGTCTGTGCTCTTCATGACCTCGCAG CTGAAGAAAGATAGAAGACTGGCGTATTGTAATAAGAATTTAGGTAGAGATAAGCAGTTT ATAGTGACGTTGCCGGACGAGGAGCGGGTGGCGTGGATGTGGGCGCTGTTCATCGCATTT ATGGTCCCCGAGATCGGGACCCTTATCAGATCTGTCCGGATATGCTTCTTCAAGTCCTCC AGAACTCCAAGCAGCGCTCAATTTATTGTGATTTTTGTATCGGAATCTCTCCACACCATC GGATTGGCGCTTTTGATGTTCAAAGTGTTGCCAGAAATCGACGTGGTCAAAGGAGCTATG ATAACGAATTGCCTCTGCATCATTCCAGCCATTCTGGGGCTATTGTCTAGAAACTCAAGG GACTCGAAAAGGTTCATGAAAGTTATAGTAGACATGGCTGCGATTGGGGCTCAAGTCACA GGATTCATATTATGGCCACTGCTGGAGAATAAGCCGGTCTTATGGCTGATACCGATCTCG
.0 TCAATCTGCATATCACTAGGCTGGTGGGAGAACTATGTCACTCGGCAGAGTCCAATCGGT
ATAATCAAGAGCCTCGGCCGCCTCAAGGAGGAGCTGAACCACACGCGCTACTACACGTAC CGCTTCATCTCCGTGTGGAAGATCCTGCTGTTCCTCATGTGCATCCTCACCAGCATCTGG CTGGACGGCGACGAGCCCGGCATGTTCTTCCAGCTCTTCAGCGAGGGGTTCGGACCGCAT AACATTGTTGTCGAAGAGATCCAACTCCAGACGGGAGGCACAATGATCCCGGACTTAGCC AACGCCACACTAACCGGAGACTCAGTGGAGGTGGCAGCGGCCTACAACTCTGCCGTCTAC GTCATCCTCATACAAGTGTTTGCCGCTTACTTCTGCTACATATTCGGGAAGTTCGCCAGC AAGATCCTGATCCAAGGGTTCAGTTACGCCTTCCCGATCAACTTGGTCATACCGCTGGTC GTGAACTTCTTGATTGCTGCTTGCGGTATCCGGAATGGTGATACGTGCTGGTTCCATGGG ACTATTCCGGATTATCTGTTCTTTGAGAGCCCACCAGTGTACTCACTAAGCGACTTCATA TCCCGCCAAATGGCATGGGTTTGGCTGCTATGGCTTCTGTCTCAGACGTGGATCACCATC CACATCTGGACGCCCAAGGCCGAGCGTCTGGCGTCCACGGAGAAACTGTTCGTACTGCCC ATGTATAACGGACTGCTCATCGACCAAAGCATGGCGCTAAATCGTAAGAGAGATGACCAG AAGGATGTTAAGACTGAGGATCTGGCCGAAATCGAAAAGGAGAAGGGCGACGAGTACTAC GAAACTATTTCTGTGCACACCGACAACACTGGGTCCTCTCCCAGAGCCGTAAAATCTTCC GATCAGATCACAAGAATCTACGCATGCGCGACGATGTGGCACGAGACGAAGGACGAGATG ATGGAGTTCCTCAAGTCCATCCTGCGGCCGGACGAGGACCAGTGCGCGCGCCGCGTCGCG CAGAAGTACCTCAGAGTCGTGGACCCCGACTACTACGAGTTTGAAACCCACATATTCTTG GACGACGCTTTCGAAATATCGGACCACAGTGACGACGATTCCCAGGTGAATCGATTCGTG AAACTGTTGGTGGACACGATTGACGAGGCTGCGTCAGAGGTGCACCAGACTAATATTCGT ATGAGGCCGCCGAAGAAATTACCTGCCCCGTACGGGGGACGGCTGACCTGGGTGCTGCCT GGGAAGACCAAGATGATCTGCCACTTGAAGGACAAGGCCAAGATTCGACACAGGAAGCGA TGGTCTCAGGTGATGTACATGTACTACCTGCTCGGCCACCGTCTCATGGAGCTGCCCATC TCCGTGGACCGCAAGGAGGTGATGGCTGAGAACACGTACCTCCTGACACTGGACGGAGAC ATCGACTTCCAACCGCACGCTGTCAGGCTGCTGATTGATTTGATGAAGAAGAACAAGAAC CTGGGCGCTGCTTGCGGACGCATCCATCCTGTTGGCTCTGGGCCAATGGTGTGGTACCAG ATGTTCGAGTACGCGATCGGTCATTGGCTGCAGAAGGCGACGGAACACATGATTGGCTGC GTGCTGTGTAGCCCCGGATGCTTCTCGCTCTTCAGAGGGAAGGCTCTCATGGACGACAAC GTCATGAAGAAATACACGCTGCGATCCGACGAGGCTAGGCATTACGTGCAGTACGATCAA GGGGAGGATCGTTGGTTATGCACATTGCTGTTACAACGAGGATACCGAGTAGAGTACTCA GCCGCCTCCGACGCCTACACGCACTGCCCTGAAGGTTTCAGCGAGTTCTATAACCAGCGT CGTCGCTGGGTACCCTCCACTATCGCCAACATCATGGACTTGCTTGCCGACTACAAACAT ACCATCAAAATCAACGACAATATCTCCACACCGTACATCGCTTACCAGATGATGTTGATT
.0 GGCGGTACGATCTTGGGCCCCGGAACTATATTCCTTATGTTGGTGGGAGCCTTCGTGGCT
GCGTTTAGAATCGACAACTGGACTTCATTCGAATACAACCTCTACCCGATATTGATCTTC ATGTTCGTTTGTTTCACGATGAAATCTGAGATACAGTTACTGGTGGCACAAATACTCTCT ACGGCATATGCAATGATAATGATGGCGGTAATCGTGGGTACAGCTTTACAATTGGGCGAG GACGGAATAGGTTCGCCATCGGCTATCTTCTTGATATCACTGTCAAGTTCATTCTTCATA GCCGCTTGTTTGCATCCTCAAGAGTTTTGGTGTATCGTCCCCGGTATCATTTACCTTCTG TCTATTCCTTCTATGTATCTCCTGTTGATTTTGTATTCGACTATAAATCTTAACGTCGTA TCTTGGGGTACCCGAGAGGTGCAGGTTAAGAAAACTAAGAAGGAAATCGAGCAAGAAAAG AAAGAAGCGGAAGACGCAAAGAAGAGTGCGAAACAGAAGTCTTTACTCGGGTTCTTGCAA GGAGCAAACCAGAATGAGGATGAAGGGTCAATAGAGTTCTCATTCGCGGGTCTATTCAAG TGCATGTTGTGCACACACCCTAAAGGCAACGAGGAAAAGGTGCAACTGTTGCATATCGCA TCTACACTTGACAAGCTCGAGAAGAAACTGGAAACTGTTGAAAAGACCCTCGACCCTCAC GGCCTCCACAGAGGTAGGAAGCTGTCGATAGGCCACCGCGGCAGTACCAACGGAGACCAC GGGCTGGACGCCCTGGCTGAAGACAATGAGGACCACAACCTCGACTCTGACACCGACACT CTATCCACGGCACCTAGAGAACAAAGAGACGAATTAATAAATCCATACTGGATTGAGGAC CCAGAATTAAAGAAGGGAGAGGTAGACTTCTTGAGTCAGTCCGAGATTCACTTCTGGAAG GATCTGATTGATAAGTATCTGTACCCGATCGATGCCAATAAGGAGGAGCAGGCCCGTATC TCGCACGACCTGAAAGAGCTGCGAAACTCATCCGTCTTTTCCTTCTTTATGATCAATGCC CTCTTTGTTCTCATCGTATTCTTGCTGCAACTGAACAAGGACAACCTCCACATAAAGTGG CCCTTCGGAGTCAAAACTAACATTACGTATGATGAGGTGACGCAAGAGGTGCTGATCTCC AAGGATACCTGCAACTAGAGCCTATTGGTCTGGTGTTCGTGTTCTTTTTCGCATTGATTT TAGTCATCCAGTTCACTGCCATGTTGTTCCATCGATTCGGAACTTTGTCGCATATATTAT CGTCTACGGAACTGAACTGGTTCTGCAATAAGAAGGCGGAAGACTTATCTCAAGACGCAC TGCTAGATAAGAATGCGATAGCAATAGTGAAGGATCTCCAGAAACTAAACGGGCTCGATG ACGGGTATGACAATGACTCGGGGTCGGGCCCGCACAATGTGGGAAGGAGAAAGACGATAC ACAACCTGGAGAAAGCGAGACAGAAGAAGAGGAACATAGGAACGCTCGACGTCGCTTTCA AGAAGCGATTCTTCAACATGAACGCTAATGAAGGACCAGGAACACCAGTTCTGAACCGCA AGATGACGTTGCGAAGAGAGACGTTGAAGGCGTTGGAAACGAGGAGGAATTCTGTGATGG CCGAACGAAGGAAGTCGCAAATGCAAACACTTGGAGCTAACAACGAATATGGAGTCACTG GAATCTTAAACAACAACCCAGCGGTGATGCCGCGCCACCGGCCGTCGACAGCCAACATTT CGGTCAAGGACGTCTTCGCGGAACCCAACGGGGGACAAGTGAACCGAGGGTACGAGACCA CGCACGGCGACGAGGGAGACGGCAACTCCATCAGACTGCAGCCGAGAACCAACCAGGTC
TCCTTCCAGGGGAGATACCAATAA ~0
[0266] > P. xylostella- Beta tubulin (PxPTUB); KX420688.1 (SEQ ID NO: 88) GCGGCAGCCAGCAGTACCGCGCGCTGACCGTGCCCGAGCTCACACAGCAGATGTTCGACG
CCAAGAACATGATGGCGGCTTGCGACCCGCGCCACGGCCGCTACCTCACCGTGGCCGCCA TCTTCCGCGGACGCATGTCCATGAAGGAGGTCGACGAGCAAATGTTGAACATCCAGAACA AGAACAGCAGCTACTTCGTCGAATGGATCCCGAACAACGTCAAAACGGCCGTGTGCGACA TACCGCCTCGTGGACTGAAGATGTCTGCCACCTTTATCGGGAACACGACAGCAATCCAAG AGCTCTTCAAGAGGATTTCTGAGCAGTTCACTGCTATGTTCAGGAGGGAAGCGTTCCTCC ACTGGTATACTGGTGAAGGCATGGACGAGATGGAGTTCACAGAGGCGGAGAGCAACATGA ACGACCTGGTCTCCGAGTACCAGCAGTACCAGGACGCCACGGCTGAAGACGAGGGAGAAT TCGACGAGGATATTGAAGACGAGTGA
[0267] > S. frugiperda-Peptidoglycan recognition protein 1 (SfPGRP1); repc7951 (SEQ ID NO: 89) GCATTAACATCTCAGGATTTGATTCGAAAGTAAGGTCTGAATTTAGTCCTTACATTTACT
TAAAATTAGGTCCTTTTTGGTTTGTACTGAAATGAAAGTTTTTCTGTTCTTGGTCTTAAT TGTGAAGATAATGGCTGAGGCAAAAGGAGATTGTGATGTGATCCCGATTACGCAGTGGGG AGATTCACCTCTTAAAAGGGAGGATYCTCTTCCAAATCCAGTGAATATTGTTGTCGTCCA AYACRCTGTGGTACCGGAGTGTAACAATGATGAAGAGTGTGAGAAAGCAGCCAMTGGAAT CAGGAGCTACCACATTAACAAACGTGGATTCACTGATATAGGACAATCGTTCCTGATTGG TGGAAACGGGAGAGTTTATGAAGGAGCCGGCTGGCATCACGTTGGGGCCCATACTTTGGG ATACAATGCAAGATCTGTGGGGATCTCCTTCATTGGCGATTTTAGAACAAAATTACCAAC ACCCGAAGCACTGAAAGCCTTCAACAGTCTCCTGGAATGTGGAGTCACGAACAATTATCT GTCAAAGGACTATCACCTGGTGGCCCATAGTCAGCTCTCTATGACTGACAGTCCYGGAGA CATGYTGAGGAAGCAGGTGGAATCGTGGCCTCMTTGGCTGGATAATGCCAAAGACATACT TAAGTAGAAPAAGACTAAACGCCGTACTTTGAGCCATTTAATGGTTACTTAACCCAGTCC TTAGCAATTTGATACAAGGCCAATGTCTCTAAGGGCGGCAGTAAAGGTCAAAACACATTT AATGAGTGTGTTTAAGATTTTGCTAGTGAAAATTGTTTTGAAGTACGTATTTGATGTAAG TGATGATATCAGTACCCTTAGTATGAGTTTGCTTTACGTTCCACGAGATGGAAACGAGAG CGCGTTCGGCGCTCTGATTGGTTCGTTCATTCATGCCGGCCAATCATAGCGCCGAATGGG CTCTCATTTCGTTTTCGTTCAACGTAAAGTAAATTCGTACTAAGGGTACTGACTTTAAAA
.0 TAAGTTACCAAAAAGAGTATTACCTATTTACATTATTTTATTTATTTTAGGTGTATTGTA
ATTCAAGTATTAAATTAATTAGTGTAGATTAATKSCATGCATTTTATATTTGATTTCATT GAATAA
[0268] S. frugiperda-Attacin (SfAtta); repc9395 (SEQ ID NO: 90) ATCATTCCAGATCCTCTCTCATACATCCAACACTTGAAGCAAACCAATCCACATACATTA
TAGCAACATGTTCGCTCTCAAGTTGGTACTAGCTGCAGTGCTGGTGGTCGCAAGCGCCAG ACATCTACCACAGGACCACTCAACGTACGACCAAGTACAACTCCTCGGGTTCGACGAAGA TGGACGACCAGTGTTTGAGCACGAAGACTTACTCCCAGAACTAGAGGAGTCCTACCAGCC AGAGCACCTGGCGAGGACTCGCAGACAGGCGCAGGGCAGCGTCACCCTCAACTCCGACGG CGGCATAGGCCTGGGCGCTAAGATCCCGCTCGCACACAACGACAAGAATGTGGTGAGCGC CATCGGCTCCATGGACTTCAACAACAAGTTGCAGCCTGCTTCCAAGGGCTTCGGTCTGGC TCTGGACAACGTCAACGGGCACGGACTGACGGTGATGAAGGAAAGTATCCCCGGGTTCGG GGACAGGCTGTCGGGCGCTGGCAAGCTGAACGTGTTCCACAACGACAACCACAACGTGGC CGTGACCGGCTCTCTCGCCAGGAACATGCCCAGCATCCCGAACGTGCCCAACTTCAACAC GTACGGCGGGGGCGTCGACTACATGTACAAGAACAAGGTGGGAGCGTCTCTGGGCATGGC CAGTACTCCGTTCTTGGACCGCAAGGACTACTCCGCGATGGGCAACCTGAACCTGTTCCG CAGCCCGACCACTACCGTGGACTTCAGCGGCGGCTTTAAGAAGTTCGAATCTCCCTTCAT GAGCAGCGGCTGGAAGCCTAACTTCGGCCTTACTTTCGGCAGATCTTTCTAGATATATTT TGTAATCTAAATTTAACTTTAACTTTGTTGTATAATATTTTGTCGAATTAAGATCAGTAT TGTTCATACTAATATTATATTATCAGTGTTTCTTATAAATTAA
[0269] S. frugiperda-CtypeLectinl5 (SfCTL15); Joint2_repc488 (SEQ ID NO: 91) AGTTTTCGTGCTTAGCACACAGAGCGCCGAACGCGCTCTCGTTTCAATTACTTTAAACGT
AAAGCAAACTCGCACTAAGGTTACTGTGACCTTTGTAAACATGTACAATGCAATACTATG TTGTTATTTTCGATACAATCATATAAAAGACAACGTTTAAAAAAAAAGCATAGCACAGAC CACAGTACAAGTATCGAGACCAAACTTTTGTTAAAACTTATTTCGTTCTGTGCAGCAACT CTAATGCGAATAATGTGCAAATTATAATAAATATGTTTTGTATATAAACATGTGTTATCT CAAGTTCAAGACATTCCGGCTCCTACAGTCAACAGATACAGTGCACAAAAATGTTAAAAA TATATTTAATTTGTTTGTCTTACTTGTTCATGTTAAACTTAGACAGGGCACAATGCACCC CCCAGTATTGGTTCAACATGGATGCGAATGGTTGGCTGAAAGTGCACACGATACCCGCCA CGTGGGAGGAAGCATTCCTTCGGTGTCACTATGAAGGTGCGGTACTTGCGTCCCCCTTGA
.0 CGCAACAACTGAGTAAGGCTCTTMAAAACAAGTTTGCAGKCWTCGGTAACCCATCAATCC
ACTTGGGGACACATGATTTGTACTCCASTGGTTACTACTTTTCTGTAGAAGGAGTACCAA TGGACAGCTTGGTGCTGAAATGGAGTAATATCAGAGGTACGGGCGAYTGCCTGGCGATGT CACGCGACGGGGAAGCATTTTTCACGAAATGCGAGCAACCTCGACCCTACATCTGCTACA AGAAGCTGGACAATCTGACGATGAACATCTGTGGAACATTYGATGACGCKTATCAATTCT ACGATAAGACTGGCAGCTGCTATAAGAGACACAACGYCTATCAGACATGGCCTGACGCGT TCAAGATATGCGCTGCCGAGGGAGGSTACCTGGTGATCCTGAACGATGACACAGAGGCCG CCATCATCAAGGATATGTTCCCCGTACGGCCAGGAAAGCCCAACGAATGGGAGAACTTCC ACGTGGGKCTGCGGGCATGGGGACCGGAACGTACTTGGATCACTATTCATGGAGAAAAAA TTGATGATGTATTCCATAATTGGAACCCGGGACAGCCGGACAACTACAAGGGAGTCCAGG ACACTGGCGCTTTCCTTAGARMAGGCACTTTAGACGATCATGCCGCTGGTGACAAATGTA TGTTTGTCTGTGAGAAGGATCCTAAAGTAAAACGCTTCGAAGAGYTACCCGAAGGATTGG CMGAAGTTTTAGGACAATAGGCCGCTAGTCAAATATTGTTCATATACCTWAGTTTTAATT ATGTAAAAATAATARTCGATTGCAGAATTTAATAAAATTTAAACTAAAAAAAAAAAAAAA AAAAAGGTMAAAACATGTC
[0270] S. frugiperda-Galectin4 (SfGIc4); repc2653 (SEQ ID NO: 92) GCCTTGTAGTTCGACAGTCAAATCCAACGGGTGTCAGAAACAATTTCCGTTTTTCCGGCT
ATTGATCGTCATATAAATAACTCCGAAGGAAAAATGACTACAATCGACAATCCGACAGTA CCGTTCACGAGACCCATTCCTGGGCATCYGCTCCCCGGCCGCAAAATGGTCGTTAAGGGT GCAATCTCTCCCAGATCAGATAGGTTCTCAATAAACTTGAAATGTGGTAGCGAGGACATC GCTTTCCACTTCAACCCTCGCTTCAGTGAGCAGAAAATAGTTCGCAACTCTTATATTTCT GGCAAGTGGGGTCATGAGGAGATCAGTGGAGGCATGCCGTTGGTAAGGGGAGAGCATTTT GAAGCGCAATTTGAATGCAATGAAGATAATTTTTCGGTGGAGTTGAACGGGAAACATTTC TGCAATTACTCTTATCGCATCCCAATCCATAAGATCACCCACGTCAACGTGGACGGTGAC GTCACGATAAGTCAGATCACCTTCGTGGACGCCTGAGCCATCGCCCGATCTTACAATGGT ACTTTTTTTTTACTTATGAACTAATGCTGGAAACCAAACTTCTTTATTCAAATATTTTTC TTTTGTCTTTATGACTATCAGTGTTTTTATTGCAATAAAAAG
[0271] S. frugiperda-Lysozyme (SfLys); repc18992 (SEQ ID NO: 93) ATCAGTGTGGTGTCTTCAACCCAAGTGCATTGTATTGCTTGGTAAATAATAACTGASAAC
AAAAATCTTGGTTATTGCTTGAACAGGTTTCGCCTATCAATTAGCCGAATATTGTTACCC T TGACACGCAATAATTGGTTACTCGATTAAAGCTTGAAGAGGTAAAGTGCCGAAATGACG CGCGCCATTCTGTTTGTTGTTGTTTTATGCTTTATTGCAAAATGCTATGGAAAAACATTC ACKGAATGTGAATTAGTTCAGGAGCTAAGAAGGCAAGGATTTCCTGAACATGAGCTTAAA GATTGGGTRTGYCTGATCGAAGCGGAGAGTTCCAAACGAACCAACGCCATTGGTAACGGC AATTCAGATGGCTCTCTCGACTATGGCTTATTCCAAATCAAATAACCGCTACTGGTGCAG CGAYGGTGACCATCCAGGCAAGGGATGCAACGATACCGCTAGTAAAGATCTGTTGCTGGA TGACATCACAATAGCGTCTCAGTGCGCTAAGACCATTTTCGGTGTCCACGGATTTAACGC CTGGGTCGCATGGGTGAACAAATGCAAAGGAAGGACCTTACCYAACCTTCACTGTTAGTT ATTTATTGAGAAAATGTAACTAAGGTATATGGTTACTTTGTACCTAAATATAGGTATTAG GCATATATTGTCCACTCTCATCAAATTTTTACTTTTATATCAGT
[0272] S. frugiperda-Hemolymph proteinase 10 (SfHP10); c12881 (SEQ ID NO: 94) ATTCGCTGTACGCACCTCGGATTGTGCGGTCAACTWTACAAGGCTCGTACCTTGCAGTTG
AACCGACAATCTATTCCTAAAGCCTTTTTAAGGTCAGGAAAAATAGTTCCTACATCTAAA TGCAGTAGAATTTGCGAAACGAATTTAAATAAAAATGGCGTCGATTGTGTTTGTGATTTT GTGTGTTACCGTCGCTGCGGTGAAAAGCGCGATTTTAAACCCGTGGAGTAAAGTTGAGGC CAACAAATGTGGTGTAGAAGCCAGTACTAACTTGGTCCATCACAATCCATGGTTGGTCTA CATCGAGTATTGGCGTGGAAACTCAGATACTGAGATCCGATGCGCCGGTACTTTAATCGA CAGCAAACATGTCGTCACAGCTGCCCACTGCGTTAGGACTCTGAAGTTTAGTCATTTGAT CGCCCGTCTTGGCGAATACGACGTAAATTCTAAGGAGGACTGCGTTCAGGGCGTGTGTGC CGATCCCATCGTCAGAATCAAGGTGGCTGAGATCATCGTGCATCCTAACTACAGCAACCG GGAACATGACATTGCAATCTTAAGGCTGGAGGAGGAAGCTCCTTATACCGATTTCACTCC GGCCCATCTGTCTGCCTTCTGGTGATCTCGCGGAAGACACCCAGTTCTTAGCAGCCGGCT GGGGTGARATCCCCACGAAAGGCTTCTTCAGCCACGTGAAGAAAATCGTCCCCTACGTAC TGGAATCGAGAGAGATGCCAAAAGGTGTACCAGTACAATTATATCCCGGAGAACGTGATC TGTGCCGGT
[0273] S. frugiperda- Trypsin like serine protease (SfTSP); repc48453 (SEQ ID NO: 95) o CAAGTAGCAACAAAATGCGTGTCCTCGCTTGCTTGGCCCTTCTCTTAGCTGTGGTAGCAG
CCGTCCCCTCCAATCCCCAGAGGATTGTGGGTGGTTCGGTCACCACCATTGACCGGTACC CCACCATTGCATCCCTGCTGTACTCGTGGAACTTGAGTTCCTACTGGCAGGCGTGCGGTG GTTCCATCTTGAACAACCGTGCCATCCTTACTGCTGCCCACTGCACAGTTGGTGACGCCG CCAACAGATGGAGAATCCGTCTTGGCTCCACCTGGGCCAACAGCGGTGGTGTCGTTCACA ACGTCAACACTAACATCGTCCACCCCTCATACAACTCTGCAACTTTGAACAACGACATCG CTATCCTCCGCTCCGCCACCACCTTCTCCTTCAACAACAATGTTCAGGCTGCCTCCATTG CTGGTGCCAACTACTTGCCCGGTGACAACACCGCCGCCTGGGCCGCTGGATGGGGAACTA CCTCCGCTGGTGGCTCTAGCTCTGAGCAGCTCCATCACGTTGAGCTGCGCATCATCAACC AGGCTACTTGCAAAAACAATTACGCTACCCGCGGTATCACCATCACCTACAACATGTTGT GCTCTGGCTGGCCCACCGGTGGTCGCGACCAGTGCCAGGGTGACTCTGGTGGTCCTCTCT ACCACAACGGCATCGTTGTTGGTGTCTGCTCTTTCGGTATTGGCTGTGCTCAGGCTGCCT TCCCCGGTGTCAACGCTCGTGTATCCCGCTACACTGCCTGGATCTCTTCCAACGCATAAG ATGTTACTTGGTGCTAATAATATTTTTTTGTAATAAAATGTTACTTTTATCCTCC
[0274] S. frugiperda- C Type Lectin 6 (SfCTL6); Joint2_ repc448 (SEQ ID NO: 96) AACAGTTTTCTATTGGCAGTCAAAGACTTCAGTCGAAAAATAATCCTCATCAGAGTCGTG
AAGCAAGGTGCCCAAAATATAATGTAACCTAGATACCTATTAATAAATTATTTGTCAACC AAAACGTTACGTTCAAAGTCCTTAAAATCAAAATATCTTATGATTAGTTTTGATTTAAAA ATAGAGGTTCGAAATCGCCAACCCAAATAGGTTTAGTTTACGATTCAGGAAAAATCCTAA CGTAGGGAAACATTATTTTACAAGACTTTTGGCTTAAAAACTTTGAGAACCAATGTCAAA TTTGATAATAACTAATGAGGTATAAAAGCTTGATCCTATTAGGACTTATTTTCATAACAC CATCGAGTTTGTATTTAATRRAGACGTGKGTTAACTAACAACATGAAGACCGGTGTAAAA TATTCTGTTMTTTGGATATTCTCTCTATTYTGCTATATAGAGGCAACATTTCGTTGTGAC TACACGTACAGCAAGGAAGCGAAGGGCTGGTTCAAACATGTGGTGATACCAGCTACTTGG GCTGACGCACGWCTGCACTGCACGTTGGAAGGTGCAACGCTGGCTTCTCCACTCAACCAG GCTATMAGTAATGAGATGCAGTCCMTCCTGGCPAACCTCTCGGCGCTGCAATCAGAAGTC TTCACTGGAATTCACRCGACTKTTTCACGRMRCAACTTATATCATACYATYGAAGGTATA CCTCTTAGTAAAATTCCATTAGATTGGGCAACAAATGAGCCAAATGGTGGGAGAGATGAA AACTGTATCACGTTTAACTCCGATGGCCAAGCGGCAGACAGATCCTGTAARGAGACTCGA CCTTACATCTGCTACCGACACACWACTAAAGTGACTGTGKCCAATGAATGTGGGACTGTA GATCCTGAATACAATTTGGATAAAAGAACGGGCKCYTGCTATAAGTTCCACACRGTACCT
.0 CGCACGTTCGAGCGTGCCAACTTCGCGTGTTCTGCTGAAGGTGGMCACCTTGCCATAATC
AACRGTGATRTCGAAGCTGCAGTACTGAAGGAACTCTTCGAAAGGAATCCACCTGCAAAG ATGTTCGGTACTTTCTGGAAAGACGTTGCTTTCATTGGTTTCCATGACTGGGGTGAATAT GGTGACTGGAGGACAATTCACGGTCAAACGCTAGTAGAGGCAGGATACAGCAAGTTTTCA TCTGGAGAGCCAAATAATTCCTCGACGGGAGAATTCTGTGGTTCCATCTATCGGAATGGT ATGCTCAACGACCTTTGGTGTGGGCATCCTAAACCATTCATCTGTGAGAAGGACCCGAAA TATCCAGCCGTATGTTGTGTTACAGAATCAGAACCAGAATTGGACCCTACTCACTTCTTA GAGTAATTCAAATTGGATCTTTTTTTATAATTCTACATCATTAGAAATATGATGTTTAGT TGAATCTGCTTTTGAAGATTGATT
[0275] S. frugiperda- Serine protease homolog 13 (SfSphl3); repc1904 (SEQ ID NO: 97) GAAKAGTGTCAATTTATTTAATTGTCAAAATCGCGTTGAATGTAAGATTGCTATAGAAAT
TATTTGTAAAAACTTTCTCAAAAATTTAATTCTAAAATGTAAAGGAACCTAAAATCAGAT ACCGATAACACATTTTGTAAATGGTTTAATATTGAAACGAAACTTCTAAACATTTTTGGT GAAATACACATAATAATATAACTTCTTCGAAAACTAGGTAGATAGACTTCAACTCAGTTT TTATTAGCGAGCTAGAAGATACAGAGATTATCTAAGATGGTGTGTGATGGTCCGGATCCT CCAGAGAACAATCCTCACGAAATTGTTCCACCTCCTAGGAATGAATTGATAATGAATGGT AATCAAGGGGATGACAGTGATGAAGAACATGAATATTTTGGCTATGAACCTTTGGCGCAA GGTCCAGAACTAGCAGTCTCAGATCACGATAGTGATGATGACCCAGAGAGTGCTGAAACA CCTCCAGCTGATGTTCCAAATATAGAGCCAATGGAAAATGTACTAAGCCGGGAAGTTTGG AGTGCCCCGAGRCATACAGATGCTATACAAATGGATAATGAACGGGCTCAACAGGTGATG AGAGCTATGGCAAATTTTGCTCTACCCCAGGCCTCAWTTCCAGAATGGGCTCAGAGCATC TCTGAAGAACAATGGAAGCAAACTCTGAATGAACAAATAGAAAGATTGAAAAATAAAAGA TAAACTAATTATATGTAGTATAATAATAATTAAGATTTAAGTGATAAAAATAACAATTAT AACTTATATGTTTAAAAAATGTATATAAACTGTAAACTTTAAGCTTATTCACATTACTTT AAATATAGAAAAAATATGTGTACCTATTTATTGTTTGGCTACATAATCCAATATAGATTA AATTGCAAATGTTTAATGTAATTATTTAGTTGGATCAAAAATAACAGCACTAAAGGTCAC TAGTTTAGTTCTTTCATTTCGAATGAAAAAGTAAAATGGAGAATCTGCATAGAAATTTTC AGCTGACCGTGTTAATGTTCCACTTGTTGCGGCTACTGCTTCTGTTCCAAACTCATTAAT TGTTAACTTTACATCATGAATTATGGAATCCACATGTATTCCAGGGTTGGGTAGACTGTC CAAAATGTTTGTCCCATTTCCTTCGTTTTCTGAGTATCTGTCTGATTCTCCTTCAGATTT ATTCACAAACTGATTGGTGTTATTTACCATAAGTGCGAAGTTTGCTTGGCCTGGTATAAA
.0 CATACTTTGTATCCCAATAGATTGCAAGGTGCTCTCTAGCTTTGTGGTGCTTTTAATTTT
CATCTTAGGGAAACGAATAACACAATTTTTCACATGCATTTCATTGATGGTATCATCAAT AACTTTGTTGTTAATATTTTCCATGAGTTCCAATAATGTTAATCGYTTATGAGATAATGG TTTTAAAGCGTACATAAAAGTTTCACTGTCGTTATATGGCAATGCTATCATGTGGAAATC ATGTTCCTCAGAAACCTTGTAACGAAACTCCCCAAAATTCAACATCATGTCAGCCATAAC TTCATCTGTGGCCCTTTTGAATTCCATCTTTCTTGTGAATTGCGGTGCAAAAGGCTGTTT CCATGTGCCACTAAAGTACAATGCAGTTAAAAGTACTACTTTAGTATGGGGAGGCAGAGA ATCTTTCAAAAAATCGTCTATATTTCCTTCTGTGTGCTTGCTCACCCACTCATTGATGCT GTTTTTAGATCCTTCGGTTTCTTGAAAATCAGTACGTAACACATCGCCTCCGTAAACACC ATGCAGATAACTTCTGTACAACTCTCGCAATTCTACTTGGTTGTCTACAAATATCGCGTC AGCGTACAAAGTTTTTGAAAACTCGTTCATATTTAAACTCTGTAGTAGTTCGCCGAACTG TTCGTGGTTCTTGCGGTTTCGTAGAATATCTTGGGAAAATCCGAGTGTCTCTGCGATTTC ATCATGTGTCATCCCAACACTTCCGAGTAATGTCATGGCAAGCAGACCAGCGACACCTAT TGGTGATATCACGATGTTTTCGTTTTTCTTTTCGTTCATCATTTTGACTAGTAAATTATA TCCAAAATTATTTATAGCTTTTGGTATCGAGTTATCTTTCGTATTTGTTGTTTCTAAGTT ATCATTTTCAGCAGTTGTGGTTGGTATTATCGTGCTTGAAATCTGACTGTCCTCTTCATT TTGTCCAGAATATTGAAAATATAATGTTGTTATAATTAAAAATAACAATGACTTGCTATC CATGTTGTGGTGCTTAAATAAGGTATTAAACGTAAACTTCACACTAACAGGAACAGGTTC CTATTATTTCCCACTTAAATCGTGAACTTACGGCATAGACACTATACACGTCTACTCGTT TGACCAACTCTGAGAGCAACTCAAAAACAACCTTGTGTGTGTATGAATAACGAACTAAAA T
[0276] S. frugiperda- Cecropin (SfCec); repc42380 (SEQ ID NO: 98) TTCGTGTCGTATCACTAGAGTTCGAAATACAAAATAATAATACATTTATTATTTTGCCAT
AATTAATAATAAAGTTATTTTATTTCATAATAATAATGAATTTCACAAAGATATTTTGTT TGTTTTTGTCTTGCTTTGTTTTGATGGCGACCGTGTCAGGAGCTCCTGAACCGAGGTGGA AATTCTTCAAGAAAGTGGAGAAGTTGGGCCAAAACATCCGCKATGGTATCATAAAGGCAG GACCCGCAGTGGCCGTGGTGGGATCAGCRGCAGCCATWGGAAAGTGAKCCCTACGACCTG AGACATGAAGACTAATATCCAYTAAAATAASAATATTGAGGCKTATAATATTAATTTATT RTRTTTGTAAATTAAATTATTTGTAAGATAA
[0277] S. frugiperda- Relish (SfRel); c13122 (SEQ ID NO: 99) ATAATATGAAGAGTGCTGCGACTAACGATTTAAGAATCTGTCGTATAAGTCGGTCGTCCG
.0 GAATTGCTTCTGGCGGCGAAGATGTCTTCATTCTGGTGGAAAAAGTTAATAAGAAAAACA
TAATGATTCGATTCTTCGAGTTGGATGAAAACGGAGAAAAGAGGTTGGACCAATGTTGGG CGATTTGTGCAAAGCGATGTTCATCACCAATACGCAATCGTCTTTCGTACTCCTCCATAC AAAAACCCTGAGACGCCAGTCGATGTGGAAGTGTATATCGAACTGGTTCGTCCATCGGAC GGCCGTACCAGTGAACCGAAGCAATTCAAGTACAGAGCCAACCAAGCGTACAAACAGATC AAGAAGAGGAAGACTGGATCTTCCTACTGTTCTATTGGCAGCTCATCTAGTGGATCGTTG AAAAGTGGTTGCGACATTCCCATATCTGTTGTCAATCATCAACCAGAAGAAGTTCCCATG GATCGCGAGCCACCGGTGCCGTCGTCGATGTATGTTTTACCCCAGGTGCATGATTCGACG ACACCAAACCAATGCGATCTGGCCAGTGCTCTGTACTCTGCTCCTGGTTCGGAGACCAGC CAGTCTCCTATATCGAGTCCGATGTGGAGTGAGCCTCACAGCGTGATGCTGCCGATGTCG CCCATTGCCAACCTTCAGCTCAACTCCGCAGACTTCGAACAGATCACAGTACCCACT
[0278] S. frugiperda- Toll (SfToll);joint2_ c3284 (SEQ ID NO: 100) AAAACCGAATAACCGGTATACCGGATAACTTCTTCAAGGATTTGAAGAATATAGAGCATT
TAGATTTGAGCTTCAATAAGATAACTAAATTGACCAGTGGAGTGTTGTCACCAATGAAAC AATTGAAGTATTTGAACCTGAATCGAAAYCATTTGGAAGTTTTRCCTGAATTCCTCTTCG CTGGCCTCAGRAAACTAGAPAAWGTRWCAATMAACGAAAATCTACTCACTTCCATAGATT CTTTAGCATTCCAAGGRGCWACGGCWTTGCATACAATATCTTTATACGGCAACAGATTAA CATTGAAGTCCAATGAACACATCCAAGACTATATGGACTTAGATCTCTACTCACCCTTYA ACACTTTGAGCGAATTAAAAAATTTGAACCTTAGTAAGAAYAAYATAAGCTCTATATTCG ATGATTGGMGGATWGTGYTRCTCAATYTGGAGTTGCTGGATCTATCTTATAAYCATATTG GAGAGTTATTGCCGGACACCTGYCAATTCCTAAGCAACAARRTYACAGTGGACCTGCAGC ACAATGACATAAGTACTGTTATTTTATATCCTACGTCTCATATGGACTTCACAGAACCCA GCATGCCATCAAACAATGTGTTTCTCTTAGACAACAACCCATTTAACTGTGATTGCCACA TATATAACCTAGCTATGCGTCTACAAGGCAAAAAGCTGCCCTACGAACCTACTTTCAACG TAGGCAAGGCCGAATGCAAATCCCCACGCCACTTAAGTGGGGATTTGTTGTCACATGTAT CCCCACTCGACTTATACTGCGAAGAAATGTCGCCTGATGTTCGTTTTAATTGTAGTTCGG TTAAAATGAGGCCTGCGTACAATGATTTGGCGTACGACTGTGATGATGTACCACCTTACT TCTCTGATGACATTAAAAACTACTCTTTGAAATTACGACACCCACCAAAAGACTTACGCA ACCTAACACTAAGTCTACTAAACCTAACCGGCATAGGATTACAAGAAATCCCMTTTACRC CGTCAGAATCGGTAAAAGTAATCGATTTATCGAACAACAATCTCACAGAGATACCKATMM
.0 GATTCTTAGAATCGAATACGACRTTGTWTTTATCGAATAATCCATTTGTTTGCGATTGTT
CTTCPAAAGATGATATTTTAGCTTTAMGRGCPAGTTAYAATGTGAATGATTTGGATATAG TYAGCTGTTCAAACGGCGTTTTGGTAACAGATTTAGAAATATCATCGCTATGCTATACAA GAACTTTAATAGCAGAAATAGGTGGTATAYTAATATTCTTAYTAATAATCTTAGTRTTTA TAATAACATTYATATTTCCTAGACAAATGGTACTRTATTGTGGTCATAGGCTMTTTCCKT GCTGGTATATCGACGATCCAGAAACTACGGCAAAAGAATATGACATATTCATATCTTATG CTCATAAAGACCAAAAATATAGTGAATAAGCTACTCCCGTAAACTAGAAAATGATTTCAA GTTAAAAGTCTGCGTTCATTACCGAGATTGGGAAGTCGGTGATTTCATTCCGGATCAGAT CAATCGATCAGTATCGAATTCGAGAAAAACGATTATTCTTTTATCGAATAGCTTTCTCGA TTCGACTTTCGCGAATTTGGAATTTCGTACCGCGCATAATTTAGCTTTGAAAGAGGGAAG AGAGAGGGTCATTTTAATACTTTTAGAGGACGTTAGTAAACATGAGAAGTTATCTGAAGA ATTAAAGTATCATATGAATATGAATACGTATCTTACATGGGATGATATTAGATTTGATGA AAAGTTAAAACGAAGGACGATACCACAGAAATATAATAGAAAGAAATTCGTAGCGCCGGC CATTTTAAAACCTATATTCAGACAGGCTACTGAGAATAATTTGAAAAAGGCACTCGATGT ACACTTTGAATAGTGCAGGCCAATTGGTGAATTTAGCTCAAAATAAGAAGAATATTGATA TGGTATAGCTTTTCCCAAATAAGGCTCTTTTAAACATGTTAAGCCCTCTCTCACCCGACC TCTCTGACTACCGCACTCAGAGACATTTTAATGTATWTTTTTTTTTTAATTGGGTACAAG CCCGCCACAACATCCCAGACCGCT
[0279] S. frugiperda- Beta 1, 3 glucanase recognition protein (SfpGRP2); EF641300 (SEQ ID NO: 101) ATGTGGTCGGTGTTAGCCGGAGTATTGGCGATCGCGTCGCTAGGCGCGGCTTGCACCCCC
AGTTTGACCACCGTCAGTGGTACCCACGCACCGGTCACCGTCTGCTCTGGTGCTCTGATC TTCGCTGATGGCTTCGACACTTTTGACCTCGAGAAATGGCAGCACGAGAATACTTTAGCT GGTGGCGGTAACTGGGAGTTCCAATACTACGGCAACAATCGCACCAACTCTTTCGTGCGC AGTGGAAGTTTGTTCATCCGTCCCTCTCTAACATCAGACGAGTTCGGAGAAGCTTTCCTC TCATCTGGACACTGGAACGTCGAGGGTGGTGCTCCTGCTGATAGATGCACAAATCCACAA TGGTGGGGTTGCGAGAGAACAGGCACGCCGACCAACATTTTGAACCCAATCAAGAGTGCT CGTGTCCGTACCGTCAATTCCTTCAGCTTCCGTTACGGACGCCTCGAAGTCCGCGCTAAA ATGCCCGCCGGAGATTGGATTTGGCCAGCTATCTGGTTGATGCCTGCGTACAACACTTAC GGTACTTGGCCCGCATCAGGAGAGATTGACTTAGTTGAGTCCCGAGGCAACCGTAACATG TTCCACAATGGTGTCCATATCGGTACACAGGAAGCAGGCTCGACCTTGCACTACGGACCT TACCCAGCGATGAACGGTTGGGAGCGCGCCCATTGGGTCAGAAGGAACCCTGCTGGCTAC
.0 AACAGCAACTTCCACCGTTACCAACTTGAATGGACACCAACTTACTTGCGATTCAGTATC
GACGACATGGAGCTTGGACGTGTAACCCCTGGCAATGGCGGCTTCTGGGAATACGGTGGT TTCAACAGCAACCCTAACATTGAGAACCCATGGAGATTCGGAAGCAGAATGGCGCCTTTC GATGAGAAGTTCTACCTTATCATGAATGTGGCTGTCGGTGGTACCAACGGATTCTTCCCT GATGGCGTCAGCAACCCATCACCCAAACCCTGGTGGAACGGATCACCAACCGCCCCAAGA GACTTCTGGAACGCGAGATCAGCTTGGTTGAACACCTGGAACCTGAATGTCAACGATGGA CAGGACGCATCCATGCAAGTCGACTACGTCCGCATCTGGGCTTTGTAA
[0280] S. frugiperda- c20042 (Sfc20042); Un-annotated (SEQ ID NO: 102) ATCAGTCGTCCCTCGTACATCCAACACTTCAAACCAAATATCTCCATATACATAGTAACA
ACATGTTCGCCCTAAAGTTGGTACTAGCTGCAGTGCTGGTGGTCGCAAGCGCCAGACATC TACCACAGGACCACTCAACGTACCGAACATGTACAGCTGCTCGGGTTCGACGAAGATGGA CGGCCAGTGTTTGAGCACGAAGACCTGCTCGCAGAACCAGAGGAGTTCTATCAGCCAGAG CACCTGGCGAGGACTCGCAGACAGGCACAGGGCAGCGTCACCCTCAACTCCGACGGCGGC ATGGGCCTGGGCGCTAAGATCCCGCTCGCACACAACGACAAGAATGTGGTGAGCGCTATC GGCTCCATGGACTTCAACAACAAGCTGCAGCCTGCTTCCAAGGGCTTCGGTCTGGCTCTG GACAACGTCAACGGGCACGGACTGACGGTGATGAAGGAAAGTATCCCCGGGTTCGGGGAC AGGCTGTCGGGCGCTGGCAAGCTGAACGTGTTCCACAACGACAACCACAACGTGGCCCTG ACCGGCTCTCTTGCCAGGAACATGCCCAGCATCCCGAACGTGCCCAACTTCAACACGTAC GGCGGGGGCGTCGACTACATGTACAAGAACAAGGTGGGAGCGACTCTGGGCATGGCCAGT ACTCCGTTCTTGGACCGCAAGGACTACTCCGCGATGGGCAACCTGAACCTGTTCCGCAGC CCGACCACTACCGTGGACTTCAGCGGCGGCTTCAAGAAGTTCGAATCTCCCTTCATGAGC AGCGGCTGGAAGCCTAACTTCTCCTTTAATCTTGGCAGGTCATTCTAGAAATATTTTTAA ACTCTTATTTAAAAATTAAATGTAAAAAATCCWGTTTGTTCATGATAATAAGAATAAATR ACAGTATTGTTCGTACTATTTACTATGTAATCTATAAATTGTATTAATAAATGAAAATTA A
[0281] S. frugiperda- rcl6438 (Sfrcl6438); Un-annotated (SEQ ID NO: 103) ATCAGTCGTCCCTCGTACATCCAACACTTCAAACCAAATATCTCCATATACATAGTAACA
ACATGTTCGCCCTAAAGTTGGTACTAGCTGCAGTGCTGGTGGTCGCAAGCGCCAGACATC TACCACAGGACCACTCAACGTACCGAACATGTACAGCTGCTCGGGTTCGACGAAGATGGA CGGCCAGTGTTTGAGCACGAAGACCTGCTCGCAGAACCAGAGGAGTTCTATCAGCCAGAG CACCTGGCGAGGACTCGCAGACAGGCACAGGGCAGCGTCACCCTCAACTCCGACGGCGGC
.0 A TGGGCCTGGGCGCTAAGATCCCGCTCGCACACAACGACAAGAATGTGGTGAGCGCTATC
GGCTCCATGGACTTCAACAACAAGCTGCAGCCTGCTTCCAAGGGCTTCGGTCTGGCTCTG GACAACGTCAACGGGCACGGACTGACGGTGATGAAGGAAAGTATCCCCGGGTTCGGGGAC AGGCTGTCGGGCGCTGGCAAGCTGAACGTGTTCCACAACGACAACCACAACGTGGCCCTG ACCGGCTCTCTTGCCAGGAACATGCCCAGCATCCCGAACGTGCCCAACTTCAACACGTAC GGCGGGGGCGTCGACTACATGTACAAGAACAAGGTGGGAGCGACTCTGGGCATGGCCAGT ACTCCGTTCTTGGACCGCAAGGACTACTCCGCGATGGGCAACCTGAACCTGTTCCGCAGC CCGACCACTACCGTGGACTTCAGCGGCGGCTTCAAGAAGTTCGAATCTCCCTTCATGAGC AGCGGCTGGAAGCCTAACTTCTCCTTTAATCTTGGCAGGTCATTCTAGAAATATTTTTAA ACTCTTATTTAAAAATTAAATGTAAAAAATCCWGTTTGTTCATGATAATAAGAATAAATR ACAGTATTGTTCGTACTATTTACTATGTAATCTATAAATTGTATTAATAAATGAAAATTA ACTATMTAAMWAA CATGTC
[0282] S. frugiperda-j2rc2367 (Sfrc2367); Un-annotated (SEQ ID NO: 104) ATCAGTCGTCCCTCGTACATCCAACACTTCAAACCAAATATCTCCATATACATAGTAACA
ACATGTTCGCCCTAAAGTTGGTACTAGCTGCAGTGCTGGTGGTCGCAAGCGCCAGACATC TACCACAGGACCACTCAACGTACCGAACATGTACAGCTGCTCGGGTTCGACGAAGATGGA CGGCCAGTGTTTGAGCACGAAGACCTGCTCGCAGAACCAGAGGAGTTCTATCAGCCAGAG CACCTGGCGAGGACTCGCAGACAGGCACAGGGCAGCGTCACCCTCAACTCCGACGGCGGC ATGGGCCTGGGCGCTAAGATCCCGCTCGCACACAACGACAAGAATGTGGTGAGCGCTATC GGCTCCATGGACTTCAACAACAAGCTGCAGCCTGCTTCCAAGGGCTTCGGTCTGGCTCTG GACAACGTCAACGGGCACGGACTGACGGTGATGAAGGAAAGTATCCCCGGGTTCGGGGAC AGGCTGTCGGGCGCTGGCAAGCTGAACGTGTTCCACAACGACAACCACAACGTGGCCCTG ACCGGCTCTCTTGCCAGGAACATGCCCAGCATCCCGAACGTGCCCAACTTCAACACGTAC GGCGGGGGCGTCGACTACATGTACAAGAACAAGGTGGGAGCGACTCTGGGCATGGCCAGT ACTCCGTTCTTGGACCGCAAGGACTACTCCGCGATGGGCAACCTGAACCTGTTCCGCAGC CCGACCACTACCGTGGACTTCAGCGGCGGCTTCAAGAAGTTCGAATCTCCCTTCATGAGC AGCGGCTGGAAGCCTAACTTCTCCTTTAATCTTGGCAGGTCATTCTAGAAA
[0283] S. frugiperda- Chitin synthase B (SfChsB);AY52599) (SEQ ID NO: 105) ATGGCGAGACCAAGACCTTATGGTTTTAGGGCTTTAGATGAGGAGAGTGATGACAATTCG
GAGTTGACTCCGTTGCACGATGATAATGATGACCTAGGACAAAGAACAGCTCAAGAGGCA
.0 AAAGGATGGAATCTGTTTCGAGAGATTCCGGTGAAGAAGGAGAGTGGGTCTATGGCCTCA
ACTGCCGGGATAGACTTCAGTGTAAAGATCCTTAAAGTCCTGGCGTATATTTTTATATTT GGCATAGTGCTCGGATCTGCGGTTGTGTCTAAGGGTACGCTGCTTTTTATCACATCACAA CTGAAAAAGGGCAAAGCAATCGTTCACTGTAATAGACAGTTAGAACTGGACAAGCAGTTT ATAACAATCCATTCGTTGCAAGAGCGTGTGACGTGGCTATGGGCAGCCTTCATAGCATTC AGTATTCCAGAAGTTGGCGTTTTCTTGAGATCAGTCAGAATATGCTTCTTCAAAACAGCA CCGAAGCCTTCTGTTTTACAGTTTTTGACGGCCTTCGTAGTAGACACCCTTCATACAATA GGCATTGGATTACTGGTGCTTTTCATCCTGCCAGAATTAGACGTGGTTAAAGGAACAATG CTAATGAATGCTATGTGCTTCATGCCTGGAATACTAAACGCTGTGACCAGAGACCGCACG GACTCTCGATACATGTTGAAAATGGCACTAGATGTACTAGCTATCTCCGCTCAAGCCACC GCGTTCGTCGTCTGGCCTCTGCTAAAAGGCGTTAGTATGCTCTGGACGATTCCTGTCGCA TGCGTATTCATCTCACTCGGATGGTGGGAAAATTTCGTCGGCGATATCGGAAAACAATGG CCAGTCCTGGAACCTGTACAAGAACTTCGTGACAATTTAAAGAAGACTCGTTACTACACA CAGAGGGTGTTGTCTTTGTGGAAGATATTCATATTCATGTGTTGCATCCTGATATCTTTG GCGGCACAAGATGACAGCCCGCTTTCTTTCTTCACGGAGTTTGCTACTGGATTTGGTGAG CGCTTCTACAAAGTTCATGAGGTTCGAGCGATACAGGACGAATTTGAAGGTTTTCTGGGC TACAAAATTATGGACTTATACTTCGATCAAATGCCAGCGGCATGGGCCACCCCACTGTGG GTGGTGCTGATCCAGGTCCTGGCTTCTTTAGTCTGTTTTATGGCAAGTTTGTCTGCCTGC AAGATTCTGATACAAAACTTCAGCTTTACATTTGCGTTGAGTCTTGTTGGACCTGTCACC ATCAACTTGTTGATTTGGCTTTGCGGCGAGAGGAACGCAGATCCCTGCGCATATAGTAAT ACGATACCAGATTATCTGTTCTTCGACATACCACCGGTGTATTTCCTGAAGGAGTTTGTG GTGAAAGAGATGTCGTGGATTTGGTTGCTGTGGCTGGTGTCGCAGGCGTGGGTGACGGCC CACAACTGGCGCTCCCGGGCCGAGCGTCTCGCCGCCAGCGACAAGCTCTTCAACAGGCCT TGGTACTGCAGCCCCGTCCTCGACGTCTCCATGCTGTTGAACAGAACCAAGAATGAAGAA GCGGAAATAACGATAGAGGATCTAAAAGAAACAGAGAGTGAGGGTGGGTCTATGATGAGC GGATTTGAAGCAAAGAAAGACATAAAGCCTTCTGACAACATTACGAGGATATATGTCTGC GCGACTATGTGGCACGAAACGAAAGAAGAAATGATGGACTTCTTGAAGTCTATCCTGCGT TTCGATGAGGATCAGAGCGCGCGTCGCGTCGCACAGAAGTACTTGGGCATTGTAGATCCT GATTACTATGAACTCGAAGTACACATCTTCATGGACGATGCTTTCGAAGTGTCGGACCAC AGCGCGGACGACTCGAAAGTGAATCCCTTCGTGACGTGTCTCGTGGAGACTGTCGACGAG GCTGCTTCAGAGGTCCATCTCACCAACGTGAGGTTGAGGCCACCGAAGAAATTCCCCACA CCGTACGGCGGCCGACTGGTCTGGACTCTCCCAGGAAAGAACAAAATGATATGCCACCTC AAAGACAAGTCCAAAATACGACACAGGAAAAGATGGTCTCAAGTGATGTACATGTACTAC
.0 CTATTGGGCCACCGCCTGATGGACGTGCCGATCTCAGTGGACCGCAAGGAAGTCATCGCA
GGGAACACCTACTTACTGGCTTTGGACGGCGACATTGACTTCAAACCGACAGCAGTCACG TTACTAATCGATTTGATGAAGAAGGATAAGAATTTAGGAGCAGCGTGCGGGCGCATCCAT CCTGTGGGCTCAGGCTTCATGGCATGGTATCAAATGTTCGAGTACGCTATTGGTCATTGG CTGCAAAAGGCGACTGAACACATGATTGGCTGTGTACTCTGTAGCCCTGGATGCTTCTCC CTCTTCAGAGGAAAGGCTTTGATGGACGACAACGTTATGAAGAAATATACCTTAACTTCC CACGAGGCACGACACTATGTGCAATACGATCAAGGCGAGGACCGTTGGTGCACGCTACTG CTGCAGCGCGGGTACCGCGTGGAGTACAGCGCGGTGTCGGACGCGTACACGCACTGCCCC GAGCACTTCGACGAGTTCTTCAACCAGCGCCGCCGCTGGGTGCCCTCCACGCTCGCCAAC ATCTTCGACCTGCTCGGCAGCGCCAAGCTCACCGTCAAGTCCAACGACAACATCTCCACC CTCTATATAGTCTATCAGTTCATGTTGATAGTGGGTACGGTGTTGGGTCCCGGCACGATC TTCCTGATGATGGGGGGAGCCATGAACGCCATCATTCAGATCAGCAACGCGTACGCGATG ATGTTGAACCTCGTACCACTCGTCATCTTCCTTATAGTCTGTATGACTTGTCAGTCAAAG ACGCAGCTCTTCCTCGCTAACCTCATAACATGCGCATACGCAATGGTGATGATGATCGTG ATAGTGGGGATAGTTCTGCAGATAGTGGAGGATGGATGGCTGGCTCCGTCCAGTATGTTC ACAGCTTTAATATTCGGTACATTCTTCGTCACCGCGGCACTACACCCGCAAGAGATCAAA TGTTTGTTGTTCATAGCAGTGTACTATGTAACCATCCCTAGTATGTACATGTTGTTGATC ATATACTCCATCTGTAATCTCAACAACGTATCCTGGGGTACCAGGGAGACACCGCAGAAG AAAACTGCTAAGGAAATGGAAATGGAACAGAAGGAAGCAGAAGAAGCGAAGAAAAAAATG GAGAGTCAGGGTTTGAAGAAGTTGTTTGCCAAGGGAGAAGAGAAGAGTGGTTCGTTAGAG TTCAGTGTGGCGGGCCTGTTGCGATGTATGTGCTGCACCAATCCAGAGGATCATAAGGAC GATCTCAACATGATGCAGATCTCACACGCGTTGGAGAAGATAAATAAGAGATTGGATCAA CTCGATGTCCCTCCTGAGCCGACCCACCAGCCCTCGCATCCGCACACACACGTGGAGACG GTCGGTGTTCGTGATTACGAAGACAGCGAGATTTCCACTGAAATTCCTAAGGAAGAACGA GACGACCTGATTAACCCCTACTGGATCGAGGACGTGGAACTCCAGAAGGGCGAGGTAGAC TTCCTCACCACCGCTGAGACCAACTTCTGGAAGGATGTCATCGATGAATACTTACTGCCT ATTGATGAGGACAAGCGTGAAATTGAACGTATAAGAAAAGATTTGAAGAACTTGCGAGAT AAGATGGTGTTTGCGTTCGTGATGTTGAACTCTCTGTTCGTGCTCGTCATCTTCCTGCTG CAGCTCAGCCAGGACCAGCTGCACTTCAAGTGGCCATTCGGACAGAAGTCCAGCATGGAG TACGATAATGATATGAATATGTTCATCATAACCCAAGACTACTTAACGCTGGAACCTATC GGTTTCGTGTTCCTCCTGTTCTTCGGCTCCATCATCATGATCCAGTTCACCGCCATGTTG TTCCATCGCCTGGACACGCTGGCCCATCTGCTGTCCACCACCAAGCTGGATTGGTATTTC AGTAAGAZGCCGGACGACCTATCAGACGATGCGCTAATAGACTCTTGGGCGTTGACAATA
.0 GCGAAGGATCTTCAACGTCTGAACACCGACGACTTGGATAAACGAAATAACAACGAACAC
GTGTCCAGGAGGAAGACCATATATAACTTGGAGAAAGGGAAGGAAACCAAACCGGCTGTT ATCAACCTCGATGCCAACGCCAAGAGGAGATTGACTATCCTGCAGAATGAAGACTCAGAA TTGATCTCCCGCCTGCCATCTCTGGGACCTAATTTGGCAACTCGTCGTGCCACGGTGCGT GCAATAAACACTCGACGCGCATCTGTCATGGCGGAGCGACGCAGGTCTCAGTTCCAAGCG CGACCTTCCGGGGGATCATACATGTATAATAACCCTCAAAACACGATTCAGCTGGACGAT ATGGTCGGGGGGCCGTCGACGTCGGGAGTGTACGTGAACCGAGGGTACGAGCCCGCCCTG GACAGCGACATCGAGGACACGCCCGTGCCCACCAGACGATCCGTTGTACACTTCACCGAC CATTTCGCGTGATAACCACCAAAATCTGACTAACATCTCCATATTACATTTTCTACTCTG TTACGAAACGATAAAGTTTAAAGTGTATTAAATAAATTGGACAGATTTGAGTAGGTTTCA CGTTTGTGTTTAAATAATTTATGAAAATAACATACCTATTGCTTTATGACCGCTTTAATA TTAAAAGAAACTCAATATATGCATTAA
[0284] T. castaneum- Peptidoglycan recognition protein LC (TcPGRPLC); DT786101.1 (SEQ ID NO:106) ATATAAAAAAAAAAAAATATTAGGCAATTTATTTACACAAATAGCACAATTTATCAATTC
ACAAGTTGTGTATTTTATTATAAATACTAAATCAACAATAGCGACTAACAATTAACACAA TTTTATTTCACTTCCTGTCACTATCGCGAGTAATAAATTCCCGCATCAAAATGCGGCCAA TTTTTGATCTCTTTGTAAACATTTGGCCCCGGACTTTCCGTTCTAAACGTCTGATTGTGA GCTACCAGTTTATAATCCCTGGCCAATTTTCCACTCTTTACCCCCTCATCTAGCAATTTC TTCGCCACGCTGATCATCTCCGTCGTCAAATGATCATGAAAAAAATTCCCAATAAAACTA ATCCCGATCGAATCATCCATGTGAAAATTCCTGATATCCCAGCCTCGTCCAACATACGCA TTCCCATCTCCACCAATTACGAAATTGTACCCAATATCGGGACTTTTCAAATTGCCCACA TGGTAGTCCTGCATGGACTGCACCCTTTGCGAACATGCAGGAAAGTCGCTACAAGTCGGG GTAACAGTGTGTGAAACGATGACAAAATGAGTGGGATGTGGCAGTGGTTTAGAGAAGTTT AAGGTGGCACGGCCTCCCCAAATTTTTTTCTCGATGATGGCACCGGGGCCCAAAGGAAGT CTTGGAGTCGCAGTGTTAGTGTCGGGAGTTTCGGGAGACTTTTCAGTTTTCCGTGGGAGT ATTACAACGCAAGTTGTGGTGACGAGGATTACGATTACGACTAGAGAAATGCCCAAATAT TTCACAGGTTTGGAGTATAAAAAGGAGTTTTTTTGGTTTTTCAGCGGGGGGGAGAGTCGG AAACAGGGGCTTTTCAGACTTGACGAAGTTTACCAATGGAATCAGTTGAATTAACAAATC CTTTTCCCTAAAGTCCTCTAAAAACAGATGATGGGGCCCCAT
[0285] T. castaneum- Peptidoglycan recognition protein 2 (TcPGRP2); XM_965754.3 (SEQ ID NO:107) ATGAGTGGCAGTGACCCTTTAACAAATACCCAACAATCCGATCAAGATTATTACCATCCA
CTCTGTTATTCAATTCAAGTGGACGACGAAAATGAACAATCAGCTCTCCTGCCCGCATTT CATCAAAGGAAAAGTTTGCGAGTTCAGGATAAAATCTTTATTGTATTTTTATTTTCAATT CTAATTACCGGACTAGCCATTGGCCTCTATCTCCTTGCAACTGAGGGACACGAATGGAAA GCTGCAGGAGTCTATAATATTACAGTTCGGGAACAGTGGCAAGCTCACGTCCCTTCATCA ACAATGCCAAAGTTGGAACTTCCCGTAAGAAGAGTTTTATTTCTTCCTGCAAATACCACT AGCTGCGGCAGCAAATCCCACTGTGCCAAAGTCCTCCAGGAACTACAATTACAGCATATG CTGCAGTGGAAAGAACCTGACATCTCCTACAATTTCATAATGACTGCAGATGGCAGAATT TTCGAGGGGAGAGGATGGGACTTTGAAACTTCTGTTCAAAATTGTACGGTTAATGATACT GTGACAGTTGCTTTTTTGGACGAATTAGATGCGAAAGCACCGACGTTTAGACAAGCTGAA GCGGCAAAAATGTTCCTGGAAGTGGCAGTAACAGAGGGAAAATTAGAACGGTGTTTTAAT ACCGCGGTCTGGGGAGGAAATAAATTCTTCATTGATTTGGCTCGAAATGTTCAAGACGTC TTATCGGAATGCGAGGGAATTACTTAA
[0286] T. castaneum- Beta 1-3 glucan binding protein (TcPGRP2); XM_966587.4 (SEQ ID NO: 108) ATGTGCGTTTGCAAGACTGTGCTATTAATCGTTGGGCTGGGGGGTTGTTTTGCGGGTCCG
GTCCGTAATTATGGGCCATTGAGACATTACAACGTTCCGCGACCCTCAATCCAAGCATTC AGGCCCCGTGGCTTCAAAGTGAGCATCCCTCATACTGAAGGCATCCAATTATTCGCCTTT CATGGAAATATTAATAAACCCCTGCACGGTCTCGAGGCCGGACAATTTTCCCAAGACGTC CTCCAAAGGGAGGGAGACGAGTGGGTGTTCCAAGACTCCAGTGCCAAATTAAACGTGGGA GATAAAATCTATTATTGGCTCTTTATCATTAAAGAAGACCTGGGCTACAGATACGATCAC GGCGAGTACGAAGTGAAAGTTTTAGCCACCCGTGACTTCGATTCTCCTCAAACAACCTCT GTGACGCCGAATCTTGCCCCCAATCTCGGCATTTGCGAAAAGGTGATGGTAAATCTCACG CGGAAGCTGCTCGATTTGCAGCAAGAAATTGAGTCCCTTAGGGAGACGAACGATATTTTG GAGGATATGGTTCAGAAGCACACTGATACGGCGACTACTCTCACTTTGGACGGCTTGATG ATAAAGGATGACGACGAACTTGTTTCGGTGATTCAAGCAATTATTAAAGATAAACTTGGA CTAAAAAGCAGGATTCAAAATGTGACGAGGCAGGAGAATGGAATGGTCAAGTTTCAAGTG GCGAGTTTGAGAGAAAAGTTGGAGGTTGTCAAAGCGGCCAAAAGAAAACTCAAGTCGTCG AGCTTTACGATCACGTATTAA
~0
[0287] T. castaneum- midgut protein (TcMDGP); XM_971351 (SEQ ID NO: 109) ATGTATCCGTTGAAGAGGATGCCCAGTGAAGAAATCAGTATCAGTGATCTGCCTAGCGAA
ATGAAAGAAGTTTTACTAGAAATTAGCCCGAACTTTGATGAAAATCTGAAACGGGCTTTC AGGAACGAAGGAGTGAGGCTGCAGAGAGTGCAGAACAATGGACGATTTATTCATCAGCTG GACGACGTTCTCTTATCCATAGACGATACCAAAATCGAGTTACGCAACCTGAAATTCCCC TGGATCCCCGACTTCCGCATCGTGGACTTGTCCAGCGACCTGCCCATGTCATGCCTCGAC CTAAACCTAAACCTGGGCAATTTGCGAATTGAGGGCGAGTACGAAGCCAACAACACCACA CTCAGGCGATGGCTCCCGGTATCTCACATTGGTCGAATCGTGATCGGTTTTAACAACGTC CGAGCGAACGGAAAAGTCGGACTCGTGCTCGAGCAGGATTCTTTCGTTCCGCAGAATTAT GATATTAGATATGAGCCGACGGATGTTGTTATTAGGGTTAGCTATCACGTGGATGGCGAG AATGAGGTGCAAAATGAGATTAGCAATTCAGATATTGAGGCCACGCTAGGCAAGACCGTG TGGGTGCAGTTGACTGAGATATTGTCCAACCTGTTGCATAGGCAATTGGGCGAGGTTGTA GTGGAGTTTTCCGTGACGGAACTCCTCGTCGATAGGGACGAGGAATACAGGGAATACGCC AAGGGACAAGCAGCGCGCGCCAATAAACTCCTGGACTCGCTTTTGTGCTCAGCCAAGGAC TATTTAGTCG CGAAGGACTTGAGGACGGTCAAAACGCCACCCTTCGACGTCGTCTTCAAA GGGAATOTCGGGGGTGC AGCAGGGGACCTTCAGCACGGGGGAAGGGTTCCTCAAGAC CTCGCCAC T ITTGACGCGGAGACACAGC T TTAGT TGTACAAGACAAOACAAACTCACG ATATATGGGGGGATCAGGCTGAGGGAGTTTATGGGTATGGCCAGGCAG TACGAGGAAACGGCCGTCTCAGGAGOATCAAAGGCTCGCTCTACAAGAACGAGATTTTC T AAGATCACTGTGAAAAAAGAAGGCGAGCGGTGCT CGACTCAGT TAGAT TCCGTOCCAA GT TGT TGT TGTAAAGTAA
[02881 T. castaneum- Chitin synthase 2 (TcCHS2); EFA 10719.1 (SEQ ID NO: 110) ATGGCGGCGCGTCATCGCTTTGCCACAGGGAGCCCTGAGGAAACAGAGCCCCTGTATTCG
TCGACGCAAATGCCCGAAAAAGTCCGGGAAAAATGGAACGTCTTCGACGACCCCCCAAGA GAGCCCACTTCGGTTCCGAAGTCAAAAGAACCTACATCGAGTGGGGGGTGAAGTTTTTGA AAGTTGTGACAATCATAACTGTGTTTTTTGTGGTCCTTGGTGCTGCAGTGGTTTCGAAAG GGACAACCTTGTTCATGACGTCACAAATAAAAAAGAATGTGACAAGGGCTTATTGCAACA AAAAGATAGACCGCAACCTCCAATTCGTCGTCTCCCTCCCCGAAGTGGAGCGCGTGCAAT GGATCTGGCTCCTCATTTTCGCTTACTTGATCCCCGAAGTGGGTACCTGGATCCGCGCCG
.0 TCCGCAAATGCCTCTACAAGCTCTGGAAAATGCCCTCCCTCTCCGAATTCCTCTCCCTCT
TCGGCACGGAAACGTGCCCCGCCATCGGAAGCGCAATTTTGATATTCGTCGTCCTCCCCG AGCTGGACGTCGTCAAAGGGGCGATGCTCACAAACGCGGTTTGTTTCGTGCCCGGAGTTG TGGCAATGTTCTCGCGCAAACCGTGCTCCATAAACGAGAACCTGAAAATGGGGCTGGACA TCGCCAGCATAACTGCACAGGCGTCAAGCTTCGTGGTGTGGCCCTTGGTTGAAAATAACC CGACCTTGTACCTAATCCCCGTTTCCGTGATTTTGATTTCGGTGGGTTGGTGGGAGAATT TCGTGTCGGAAACGTCCTACTTACCGTTTATCCGGGCTCTGGGCAAGAGCAAAAAGGAGT TTAAGACAAGGACGTACTTCATTTACGCGTTCGTGGCCCCGGTTAAATGCCTGGCGTTTT TTTGCACCGCTTTGGTCATTTTTTACTGCCAGGAGGGCAGTGTTGACTTTTTATTTGATA ATTTTTCAGCCGCGTTTCAGGATCATAACATTGAAATTACCGAAGTCGCGCCCGTCTTGC CGGGGAATTACGCAAATGCAGTTCGGTCGGGAGCCGAAACCCCATCCACACAAGCAGTTA CATGACGGGGATTTGGGTTTGGTTGATTAACATTTCGGCGACTTATATTTGCTACGCGTT TGGGAAATTCTCCTGCAAAGTCATGATCCAGAGCGTTAGCTTCGCTTTTCCGATCAATTT GTCGGTCCCTGTCCTCTTATCCGGACTGATCGCAATGTGTGGCATGTACTACAGGGATGA GTGTTCTTTCGCTGAGTCAATTCCTCCATATTTGTTTTTCGTTCCTCCACCTCTCATGTT CCTACAAGATTTTCTCTCGCACCAACACGCCTGGATTTGGCTGGTTTGGTTGCTGTCACA AGCCTGGATTTCGGTGCACATTTGGTCCCCAAACTGTGACAAACTTTCAAGCACCGAACA GTTGTTTATTCGGCCCATGTATGACGCGTTTTTGATCGATCAAAGCCTGGCGTTAAACCG GAGACGTGACGAAAATCCCAGAAATTACAGAAGCGACGAAGGGCCTCAAATTACAGAGCT CGAGCCGGAAACGATCGAGAGTCAGGACGCCATAACCCGGATTTACGCCTGCGGGACAAT GTGGCACGAAACTCCCGAAGAAATGATGGAATTTTTGAAATCGGTGTTCCGCTTGGACCA AGACCAGTGTTCCCACAGGATTGTGAGGGAGCATTTGGGACTCAAGCATGACAATTACTA CGAATTGGAGACTCATATATTTTTCGATGATGCGTTTATTCGGACCAGTGAAGACGATAA TGATCCCCACGTCAACGAATACGTTGAGTCACTTGCGTCCATTATCGACGAGGCTGCGAC TAAGGTTCACGGTACCACGGTGAGGGTGCGTCCGCCCAAAGTGTACCCCACGCCTTACGG CGGACGCCTGGTCTGGACGCTCCCAGGGAAAACAAAAATGATCGCCCACTTGAAGGACAA GAAGAAGATTAGGGCGAAAAAGCGCTGGTCTCAGTGCATGTACATGTACTTTTTGCTCGG ATTCAGATTGCAAGCCAACGACGAACTCTCCGCCCACAGCAAGGAAATCCGCGGCGAAAA CACCTACATCCTCGCCCTGGACGGCGACATCGATTTCCAACCCGAAGCCCTGCACCTCTT GGTGGACTACATGAAGAAGAACAAAACGTTGGGGGCGGCCTGCGGCCGCATCCACCCCAT CGGCAGCGGCGGCATGGTCTGGTACCAAATGTTCGAATACGCCGTCGGTCACTGGATGCA AAAAGCCACCGAGCACGTCATAGGCTGCGTCCTCTGCAGCCCCGGCTGTTTCTCCCTGTT CCGGGGAAAAGCCCTCATGGACAAAAGCGTCATGAAGAAGTACACCACTCGATCGACCCA
.0 AGCCAAGCACTACGTGCAGTACGATCAAGGGGAGGACCGGTGGTTGTGCACTTTGTTACT
CCAGAGGGGCTACCGTGTGGAATACTCCGCAGCCTCGGACGCTTTCACGCACTGTCCGGA AGGCTTCAACGAGTTTTACAACCAGCGGAGGCGCTGGATGCCGTCCACTATGGCCAACAT TTTGGACCTTTTGATGGATTACGAGCACACGGTCAAAATCAACGAAAATATTTCCATGCT GTACATCGGGTACCAAATTATTTTAATGATCGGTACGGTCATTGGCCCCGGTACTATTTT CCTCATGTTGGTCGGCGCCTTCGTGGCTGCCTTTGGGCTCGACCAATGGAGCAGTTTCTA CTGGAATTTACTACCAATCGCAGTTTTTATCCTAGTATGTGCCACTTGTAGCTCCGATAT CCAATTATTTTTCGCCGGCCTTATCAGCGCCATTTACGGCCTGATAATGATGGCTGTTTT CGTCGGTGTGATGCTCCAAATCAGCCAAGACGGCCCACTTGCGCCTTCCTCCCTTTTCTT CTTCTGCATGGCTGCTGAATTTATAATCGCAGCACTGCTGCATCCGCAAGAATTCAACTG TTTGAAATACGGGGTCATTTACTACGTCACGGTCCCCAGCATGTACCTCCTCCTAGTCAT CTACTCGGTCTTCAATATGAACAACGTGTCCTGGGGGACGCGCGAAGTGACAGTCGTGCC CAAGCCTGACCCCAACGCCGTCCAGAAAATCGAAGAGAAAAAACCGGAGAAGAAAGACAA AGTTTTGACGTTTCTGGGCGCGAATGCCCAGGACGACGAAGGCGGGCTTGAATTTTCGGT CAACAAACTTTTCAAATGCATGATTTGTACGTACAAGGCCGATAACAAGGAAAACGAGCA GTTGAGGAAAATACAAGAGTCGTTGAGAGACTTGAATAGGAAAATCGAGTCGCTGGAAAA AATGCAATATCCGGATTTGAGGTCTCCTGCCGTTAGCAACGTTACAACGTTCATGGAGGG CTCAAAGGCGACGGTTAAGAACAACGTGGAGGATAACTACATGGAGGCTCCGCAAGACAA TGTTTCGCAACCGTCGGATGAGGTCATGGAGAATAGTTGGTTCTACGATGGGCCTTTGAT TAGGGGGGAAGTGCATTACTTGAATAGGAATGAGGAAACGTTTTGGAATGAACTGATAGA GCAGTATTTACACCCGATTGAGGATGACAAGAAGAAGGTTTCGGCTGAATTGAAAGATTT GAGGGACAAAATGGTGTTTACTTTCCTGATGTTGAATTCGCTCTACGTTATTGTGATTTT TTTGCTCACTTTGAAGAAGGATTTGCTCCATCTGGACTGGCCGTTTGACCCCAAAGTGAA CTTCACGTATTTCGAGGACAAAAATGAGATTGGCGTTTACATAACATACCTCCAGCTGGA GCCCATCGGTTTCGTGTTCCTCATATTTTTCGCCCTGCTTATGGTGATCCAGTTCTTCGC AATGATGATCCACCGCTTCGGCACCTTCTCCCAAATCATCACAAAAACACAATTAGACTT CGATCTGTGCAGCAAACCAATCGACGAAATGACTGTGGACGAACTCAGGTCCCGCGACCC TATAAAAATAGTCGCAGATTTACAAAAACTAAAAGGTATAAACAACGAATACGAGGACCA AACGGAAGTTCCGGTCGAAATGCGAAAAACAGTAAGTAATTTGGCGCAAACGAGCGGTGG TGGTGAGAACAAGCCCATATTTTATTTGGATGAAGCGTTCCAAAGGCGAGTCACTCAAAT AGGGAGCACTTCAAGCAACAACCCGAGCATTAGCGCCTTTAGGAAGAAAAGCCTTGCTTA CGTCCAACAAAGAATGAGCATAGCCCCAAATCGGGTGTCACAAGCCCGGCCTAGTGTGCA GTTACGGTACCCCAATGGAAAAGCCAACGAAAATTTCGTTTTTGACGAAAACGGGTCAGA
.0 CGTGGAGGCATAA
[0289] > M. sexta-Peptidoglycan recognition protein 2 (MsPGRP2); GQ293365.1 (SEQ ID NO: 111) TAATCATTAGAAAAATGGCGAGCTTCGCTTTAATAGTTATCCTTAGCGTAATTGGCTTTA
TATCGGCCTATCCTAGTCCTGAAGGTTACAGTTCTGCCTTCAACTTTCCATTCGTAACCA AGGAGCAGTGGGGCGGCAGGGAGGCACGCACGTCGACGCCACTCAACCACCCAGTGCAGT TCGTGGTGATCCACCACAGTTACATTCCCGGCGTGTGCCTCAGCCGGGACGAGTGCGCGC GCAGCATGCGCTCCATGCAGAACTTCCACATGAACAGTAACGGGTGGAGTGATATTGGAT ACAACTTCGCTGTCGGCGGTGAAGGGTCGGTGTACGAGGGCCGCGGCTGGGACGCGGTCG GCGCACACGCAGCTGGCTATAACAGTAACAGTATCGGCATCGTGCTCATCGGCGATTTTG TTTCAAACCTCCCGCCGGCGGTGCAAATGCAAACCACACAAGAATTGATCGCAGCGGGCG TGCGACTCGGTTACATCAGGCCCAACTACATGCTCATCGGGCATCGTCAGGTCTCCGCCA CTGAGTGCCCAGGAACCAGACTCTTCAACGAAATCACCAACTGGAACAACTTCGTGAGG
[0290] > M. sexta-Beta-1, 3-glucan-recognition protein 2 (MsPGRP2); AY135522.1 (SEQ ID NO: 112) GAGCGTCTGTTTGTTCGCAACCATTGCGGGCTGCTTGGGCCAGCGAGGGGGTCCATACAA
GGTGCCTGATGCGAAACTCGAAGCTATCTACCCCAAAGGCTTGAGAGTCTCTGTGCCAGA TGATGGCTACTCCCTATTTGCCTTCCACGGCAAGCTCAATGAGGAGATGGAAGGTTTAGA GGCTGGCCATTGGTCCAGAGACATCACCAAAGCGAAGCAGGGCAGATGGATATTCAGAGA TAGGAATGCTGAGCTGAAGCTTGGAGACAAAATTTACTTCTGGACTTACGTTATTAAGGA TGGATTGGGATACAGGCAGGACAATGGAGAATGGACTGTTACAGAATTCGTCAATGAGAA CGGTACAGTGGTGGACACTAGTACAGCGCCGCCACCAGTAGCACCCGCCGTTTCAGAGGA AGATCAATCGCCAGGTCCTCAGTGGAGACCTTGCGAAAGATCCCTGACTGAGTCCTTGGC CCGCGAACGCGTTTGCAAAGGCAGCCTTGTCTTTAGCGAGGACTTTGATGGTTCCAGTTT GGCCGACTTGGGCAATTGGACCGCTGAAGTCAGATTCCCTGGCGAACCGGACTACCCGTA CAACTTGTACACTACGGACGGCACTGTGGGATTCGAAAGTGGGTCTCTGGTGGTGAGACC CGTCATGACCGAGTCCAAATACCACGAGGGCATCATATACGACCGCCTCGACCTTGAGAG ATGTACAGGACAGCTGGGTACGCTGGAATGCAGGCGAGAGAGCAGCGGCGGTCAGATTGT ACCACCTGTGATGACAGCTAAACTGGCCACTCGACGCAGCTTCGCGTTCAAGTTCGGCAG GATCGATATAAAGGCGAAGATGCCGCGCGGGGACTGGTTGATACCAGAACTCAACCTCGA ACCTTTAGATAACATATACGGCAACCAGCGATACGCTTCGGGTCTCATGCGGGTCGCGTT
.0 CGTGAGAGGAAACGATGTATACGCCAAGAAGCTCTACGGAGGTCCGATAATGTCCGACGC
GGACCCGTTCAGGTCCATGCTGTTGAAGGACAAGCAAGGGTTGGCCAACTGGAATAATGA TTACCACGTCTACTCGCTGCTGTGGAAGCCTAACGGTTTAGAGCTGATGGTGGACGGTGA AGTGTACGGCACCATCGACGCTGGCGATGGCTTCTACCAGATTGCGAAGAACAACCTCGT GAGCCACGCCTCGCAGTGGCTCAAGGGCACCGTCATGGCGCCGTTTGATGAAAAGTTCTT CATCACTCTGGGTCTTCGCGTGGCGGGTATCCACGACTTCACGGACGGTCCGGGCAAACC TTGGGAGAACAAGGG
[0291] > M. sexta-Relish family protein 2A (MsREL2A); HM363513.1 (SEQ ID NO: 113) ATGTCCTCTTGTCCAAGCGACTATGATCCCAGTGAATCGTCCAAATCTCCACAAAGTATT
TGGGAGTCAGGAGGATACAGTTCTCCGTCGCAACAAGTTCCTCAATTGACTTCTAACTTA ACAGAATTGTCTGTTGATCACAGCTATAGATACAATGGAAATGGACCATATCTACAGATC ACAGAGCAACCACAGAAATACTTTCGGTTCCGTTATGTTAGCGAGATGGTGGGAACACAT GGATGTTTGCTTGGCAAATCTTATACAACAAACAAAGTTAAAACTCATCCGACAGTTGAA CTCGTGAATTACACCGGTCGAGCCCTGATAAAGTGCCAACTATCGCAAAACAAGAGCGAA GACGAACACCCGCACAAACTGCTCGATGAACAAGACAGAGACATGAGCCACCACGTTCCC GAGCACGGCAGTTATAGAGTGGTATTTGCTGGTATGGGTATAATTCATGCTGCCAAAAAG GAAGTTGCGGGGTGGCTCTATAGAAAATATATACAGCAGAACAAGAATGAAAAGTTTAAT AAGAAAGAGCTCGAAGCGCATTGTGAGAGGATGTCCAAAGAGATCGATTTAAATATAGTT AGACTGAAGTTTAGCGCTCACGATATTGACACTGGCATTGAAATTTGCCGGCCAGTGTTC TCTGAACCCATTTATAATTTGAAGTGTGCGTCTACGAATGATTTGAAAATATGCCGCATA AGCCGTTGTTACGGTAGACCGAGAGGCGGCGAAGATATCTTCATATTTGTCGAAAAGGTC AACAAGAAAAACATCCAAGTTCGGTTCTTTAGACTGGAAAACGGGGAGCGCACCTGGTCA GCGATGGCGAACTTTCTGCTAAGCGATGTTCACCACCAATACGCTATCGCTTTTAGAACG CCACCGTACGTCAATCACCAAATTTCTGAAGACGTGCAAGTTTTTATAGAACTCGTACGC CCTTCAGACGGTAGGACGAGCGCTCCCATGGAGTTCACATACAAGGCTGAGCAAATCTAT AAACAGAACAAGAAACGTAAAACTACTTCGTCGTACTCGTCGCTCGACAGCTCCTCAGGT TCGGCCGGTTCAATTAAAAGCATCAGCGAACTGCCCGCGCCCGTTGTTTTTGCTGAAAAC GTAAGTTTTTTCTATGACACATTACTCATTCTTCAACCCATGACGAATCTATAA
[0292] > M. sexta-Spatzle (MsSPZ1A); GQ249944.1 (SEQ ID NO: 114) ACGAACAACCTGACAGACGGATAGCGGGACGGTCAGCACAATACGAACATTTAAGAACAA
.0 ACGAGAGGTCTCTCCCGGTCTACAGCGAGACCCAGAGGATACAAGCAGAAGAGAGAAGAA
GACACAGTTCGAGACTAGAAGAACCGAGACAACGTGCTGAGAATGGTTCATATAAGATAT TGAATAACCCTCCGAAACCCTGTATTACTAATAGGAGAAGTCAAATTGATTCGTCGAATG ATAGGGTAGTGTTCCCCGGTCCGACTTCAGAAAGGTCGTACGTACCCGAAGTGCCAGAGG AATGCAAGAAAATCGGCATATGCGACAGTATACCGAATTACCCAGAAGAACACGTAGCTA ATATTATATCTCGACTTGGAGACAAAGGAAAAGTATTACAAATAGACGAACTGGACGTAT CAGACACTCCAGATATCGCCCAGAGGTTGGGTCCGCAGGAGGACAACATGGAACTATGTA GCTTTAGAGAAAAGATTTTTTACCCCAAGGCAGCGCCAGACAAAGATGGAAATTGGTTCT TCGTTGTGAATTCAAAAGAAAACCCAGTACAGGGTTATAAAGTTGAAATTTGCGACCGTC AGCAATTACCATGCGCGGAGTTCGCGAGCTTCCAACAGGGATATGAAGCGAGGTGCATCC AGAAATACGTTCGCCGGACCATGTTGGCGTTGGATCCCAAGGGTCAGATGACCGACATGC CCCTTAAAGTGCCCAGCTGTTGCTCATGCGTGGCCAAATTGAC
[0293] > M. sexta-Toll receptor (MsTOLL); EF442782.1 (SEQ ID NO: 115) ATGCAGGCTCGGCGGTGGTGCGCGGCACTGCTATTAATGCAGATGCTGAGCTGGCTCGGA
GTCAGTGGACACTTACCGCGTCCCGAGTGCGCGCCAGCCGCAGATTGCCAACTTATACGA GACAACATAATCGATGGATATGCACAATTCTACTTCAACGTATCAGGACATGAAGTGAAA TTTGAACATTACATCGGAAACGACTTCGATGTCGAATTGTCATGCAATTACATCGCCATG GACAACGCAATGCTGCCGCGGTTCTCAACGACCTTTTCAGTCAACGTAATAGTGGTTAAA GAATGTGCTTTGCCAAGAAGTGGGTCAATCGATGCCGCTGTCGCTGCACTTAATATCAAC GTTTTGACGGAGCTGACTCTGGACAAATTCCTAGAGCCGGCGGTGATCACGCGCGCACAT CTTACCAGTTTACAACGACTAGAGAGGCTGGAGCTACACGGTAACTCAAACACAAGCCTC GCCCCCGGCGCACTGGCCGCGCTCTCCGCCGCGTCCGCACTGAAATGTCTTGTATTGCAT GCAGTACGCGTGCCCGCCGCTGACCTGGCGCGCTTGCCGTCGTCACTGCAAGAACTAGCG TTGTTGGATGTGGGCGCTGCGAGTATGCATTTAGATTCATCGGTTAATTTGACGTCACTC TTCGTAATCGATACACATTATCCTGTCGTCGTGAATGTGAGCAACGCCGTTGCGCTCAGA GACTTGCACATAAATACCCCAAGTACTGTGTTGACCGAAGACGTGCTCCCGTCGTCACTC AACTCACTTGAACTAGAGGGGTGGAACGAAACGCATCCGGTGCCTAAGACACGTTGTGTA CTACTTAAGGAACTTAATGTAATCGGCACCGACAATGATGCCTATCCGGTGACTCT
[0294] > M. sexta-Valine Rich Midgut Protein (MsVMP1); NCBI accession number not assigned as yet (SEQ ID NO: 116) .0 ACGGACCTTCCGTTGGACCNGCCATCATCGGCGCTGGAGACATCGCTGTCGGCCCTGCTA
TCGTCGACTTCCCTTTCCCCGACGGCGGTGCCGTGTCTGCCCCCGTTGAGCCTTCCCCCA TCGCCATCGGACCCGCTATCGTCGGTGAATCCCCTATCTCCGTCGGACCTGCCATCGTTG AGGCCGGAGACATCGCTGTTGGACCCGCTATCATCGACTTCCCCCTTCCCGACGGTGGCG CCGTGTCCGCCCCCGTTGAGGTTTCTCCCGTCGACTCCGTCGTCGTCGGCCCTGCCGCCG GCTCTCAGAGCTCTCCCCTCGTCCAGATCATCATCAACGTTAAGGCCCCCGCTGGTGCCG GCCCCGTTGTCGATGCCGTCGCTGACAAGCCCATGGACATCATTGATGTTATGCCCGTCG TCGACCCTGCTGATTTCGTGGACCTCACCCCCGTTGTAGAGCCTGTAGAAGTCGTCGACA TTGTCGATGTCATGCCCGTGGTTGACCCCATCAACATCATCGATGTTATGCCTGTTGTTA AGCCCGTAAACCCCCTTGCCCGTTCTT
[0295] > M.sexta-Chitin synthase 2 (MSCHS2); AY821560.1 (SEQ ID NO: 117) ATGGCCGCAACTACACCAGGTTTTAAGAAGTTAGCAGACGATTCTGAGGATTCAGATACA
GAATACACCCCGCTGTATGATGACGGTGATGAAATAGATCAAAGAACTGCACAAGAAACA AAAGGATGGAATCTATTTCGAGAGATTCCGGTGAAAAAGGAGAGTGGATCTATGGCCACA AAAAATTGGATAGAAACAAGCGTAAAAATCATAAAAGTGCTTGCCTACATATTGGTTTTT TGTGCTGTACTGGGTTCCGCAGTCATAGCTAAAGGAACTCTTCTATTTATTACGTCACAA CTGAAGAAAGACAGACAAATTACTCACTGCAATAGACGACTTGCTTTAGACCAACAGTTC ATAACGGTACACAGTTTAGAAGAAAGAATAACATGGCTATGGGCAGCACTTATTGTATTC GGTGTGCCGGAGTTAGGGGTGTTTTTGAGATCCGTCAGGATATGCTTCTTCAAAACTGCC AAGAAACCAACCAAAACACAGTTTATTATTGCTTTCATAACAGAGACACTACAAGCAATA GGAATAGCAGCACTTGTATTAATAATTCTACCAGAATTAGACGCTGTGAAAGGAGCCATG TTGATGAACGCCACGTGCGCTATCCCTGCATTGCTAAACATTTTCACGAGAGACCGAATG GATTCTAAGTTTTCTATAAAATTGATATTGGATGTATTGGCGATATCGGCACAAGCCACG GCGTTTGTTGTTTGGCCTCTTATGGAAAGAACGCCAGTTCTATGGACCATACCAGTTGCA TGTGTGTTAGTGTCTCTAGGCTTCTGGGAGAATTTTGTTGACACCTACAATAAAAGTTAT GTTTTTACGGTGCTGCAGGAACTACGCGACAACCTCAAGAGGACTCGGTACTACACTCAG CGGGTGCTATCTGTTTGGAAGATTATAGTGTTTATGGCATGCATTTTAATATCGCTGCAT ATGCAAAATGACAATCCGTTTACCTTTTTCACTCACGCCAGCAAAGCCTTTGGAGAGAGA CAGTATGTCGTTAACGAGGTTCTAATAGTAGTCCGAGATGACGAAACCATAGGCTATGAC GTCACCGGAGGTATATTCGAATTGGACGCGATATGGACCTCAGCATTGTGGGTCGCATTA ATTCAAGTGGGAGCAGCCTACTTCTGTTTCGGAAGTGGCAAGTTTGCTTGCAAAATTCTT ATACAAAATTTTAGTTTCACTTTAGCATTGACTCTCGTCGGGCCCGTGGCAATCAACCTC
.0 CTTATTGCTTTCTGCGGAATGAGAAATGCAGACCCTTGCGCTTTCCATAGAACTATACCT
GACAATTTGTTTTACGAGATACCACCTGTG
[0296] > M.sexta-Beta-fructofuranosidase 1 (MsSucl); GQ293363.1 (SEQ ID NO: 118) TTGCTGTGCGTTTTCCTTGGTAGTGTATCGTCATGTTGCGTTAATGGGCGGTACTACCCG
AGGTACCATTTGTCGCCACCGCATGGCTGGATGAACGACCCCAACGGATTCTGCTACTTC AAAGGTGAATACCATATGTTTTACCAGTACAATCCCATGTCAAGTTTGGAGGCTGGCATA GCTCATTGGGGTCATGCGAAAAGTAAAGATTTGTGCCATTGGAAACACTTAGACCCGCCA TCTATCCTGATCAGTGGTACGATCAAACGGGAGTATTTTCTGGAAGTGCGCTAGTAGAGA ATGACGTCATGTACCTTTATTATACTGGAAATGTAAATCTTACTGATGAAATGCCATTTG AGGGACAATTCCAAGCTCTTGGTATCAGTACTGACGGTGTCCACGTAGAAAAGTATAAAG ACAATCCAATAATGTACACGCCAAACCATCAACCTCACATCCGAGACCCAAAAGTTTGGG AACACGACGGCTCTTATTATATGGTCTTAGGAAACGCATATGATGATTATACAAAGGGCC AAATAGTTATGTACGAATCATCAGACAAGATCAACTGGCAAGAAGTAACTATACTATATA AATCAAATGGATCTTTCGGTTACATGTGGGAGTGTCCAGATTTATTCGAAATAGACGGCA AGTTTGTACTTCTGTTCTCTCCTCAAGGCGTGAAGTCTGTGGGCGATATGTACCAGAATC TGTATCAAGCAGGATACATCGTCGGAGAATTCGATTACGATACTCATTCATTCACAATAC TAACCGAATTCAGAGAATTGGATCACGGTCATGATTTTTACGCTACACAAACAATGAAAG ATCCTAGTGGAAGAAGAATAGTCGTTGCTTGGGCAAGT
[0297] > M. sexta- Beta-1 tubulin (MspTub); AF030547 (SEQ ID NO: 119) ATGAGGGAAATCGTGCACATCCAGGCTGGCCAATGCGGCAACCAGATCGGAGCTAAGTTC
TGGGAGATCATCTCTGACGAGCATGGCATCGACCCCACCGGCGCTTACCATGGCGACTCG GACCTGCAGCTGGAGCGCATCAACGTGTACTACAATGAGGCCTCCGGCGGCAAGTACGTG CCGCGCGCCATCCTCGTGGACCTCGAGCCCGGCACCATGGACTCTGTCCGCTCCGGACCT TTCGGACAGATCTTCCGCCCGGACAACTTCGTCTTCGGACAGTCCGGCGCCGGTAACAAC TGGGCCAAGGGACACTACACAGAGGGCGCCGAGCTTGTCGACTCGGTCTTAGACGTCGTA CGTAAGGAAGCAGAATCATGCGACTGCCTCCAGGGATTCCAACTCACACACTCGCTCGGC GGCGGTACCGGTTCCGGAATGGGCACCCTCCTTATCTCCAAAATCAGGGAAGAATACCCC GACAGAATTATGAACACATATTCAGTTGTACCATCACCCAAAGTGTCTGATACAGTAGTA GAACCTTACAATGCAACACTGTCAGTCCACCAACTCGTAGAAAACACCGACGAAACCTAC TGTATCGACAATGAGGCTCTCTATGACATCTGCTTCCGCACGCTCAAACTTTCCACACCC
.0 ACATATGGCGACCTTAACCACCTGGTGTCGCTCACAATGTCCGGCGTGACCACCTGCCTC
AGGTTCCCCGGTCAGCTGAATGCGGATCTCCGCAAGCTGGCGGTGAACATGGTGCCCTTC CCGCGTCTGCACTTCTTCATGCCGGGCTTCGCTCCGCTCACGTCGCGCGGCAGCCAGCAG TACCGCGCCCTCACCGTGCCCGAACTCACCCAGCAGATGTTCGACGCTAAGAACATGATG GCGGCGTGCGACCCGCGTCACGGCCGCTACCTCACCGTCGCCGCCATCTTCCGTGGTCGC ATGTCCATGAAGGAGGTCGACGAGCAGATGCTCAACATCCAGAACAAGAACTCGTCGTAC TTCGTTGAATGGATCCCCAACAACGTGAAGACCGCCGTGTGCGACATCCCGCCCCGTGGT CTCAAGATGTCGGCCACTTTCATCGGCAACTCCACCGCTATCCAGGAGCTGTTCAAGCGC ATCTCTGAACAGTTCACCGCTATGTTCAGGCGCAAGGCTTTCTTGCATTGGTACACCGGC GAGGGCATGGACGAGATGGAGTTCACCGAGGCCGAGAGCAACATGAACGACCTGGTGTCC GAGTACCAACAGTACCAGGAGGCCACCGCCGACGAGGACGCCGAGTTCGACGAGGAGCAA GAGCAGGAGATCG
[0298] > P. xylostella- Peptidoglycan recognition protein 2 (PxPGRP2); ACB32179.1(SEQ ID NO: 120) CCCGATACAGTTGGAGTACCTGCCCCGGCCCCTGGGGCTGGTGGTGGTCCAGCACACCGC
CACCCCCGCGTGTGACACTGACGCCGCGTGTGTGGAGCTGGTGCAGAACATACAGACCAA TCATATGGATGTGCTGAAGTTTTGGGATATTGGACCGAACTTCCTGATTGGTGGGAACGG CAAGGTGTACGAGGGCCCTGGTTGGCTGCACGTCGGCGCCCACACTTACGGCTACAACAG GAAGTCTATCGGGATCTCTTTCATTAGGAATTTTAATGCTAAGACCCCAACAAAAGCAGC GTTGAATGCGGCTGAAGCATTGCTGAAGTGTGGAGTGAGAGAAGGACACCTGTCTCACTC ATACGCAGTGGTCGGCCATAGACAACTGATCGCAACAGAGAGCCCAGGCAGGAAACTGTA CCAAATCATCAGGCGCTGG
[0299] > P. xylostella- Immune Deficiency Protein (PxIMD); Px003008 (SEQ ID NO: 121) TCCCGAAGCCACCTAGAGAACCGGTAGAGACTACAGAGACATCACAAAATAATACCGAAA
ATCAACCTTACAATGTCGAAGAAGAGGAAATACCCGAACCAGAAAAGCCTAAGAAAGAAA AAAAGAATCCCAAGCCTACCAAAAAAACTTTCTTTAATCGTGACAAAACTAACAAACACG ACGATACCCGCAAACATACAAAATCCGGAAAGGACCAGACATCAATTAATACTCAAGGTA ACTTGAAAATTATACTTCCTGTAACTTAAACGGCCCGTTGACCCCAGTTTACCTTTCGCC TTTCTTGATATATTTTTGTAATCCAGCCTTACTTTGGTAATACATACTTGCCCCACTTGT
.0 ATTTAGTTAATGGTGGCACTAGCTAGATAGTAATGTTAAATGATGATAAGCAGTAGTGAT
TCATCATTCAAATGTATCATTGTCCTTTAATGTTAAGCGCAAATAGATTTTCATTGTTCT CCCATGTGCTTCATGTTTTATGTATTTATAGGTAGGTACTTAATGTTTTATAAATATTTT TTTGTTAATTGGGAATCCCCAGTCCCCATTGTCTGGACCAGTTTATATATAATTGAACTA ACAAGAGTGTGCTTTAAATACTATTCTCTGCAATTATGATAATTAAACAACATGAATTTC TCTTCACTTCCCTTCTCTTATTTAAATAATATTGTAGGAAACTGTAATAACTAATACAAG ATTATAAATTTCATTCTAGCAACTGGTGATGTAATCCATGTGGTAAATTCCAAAGATGTG CAGGTCGGCCATCAGTATGTGTACAACATGGGAACTCCCGGAGCTAACTCACAGAAGAAT AACCCATTTGATGATGAAGAAACAGTAGAAAAGACAAATCTAATAACTCTGGTCATGGAA GCAAAAATTATGGTAATAACACATTTTTAACTAGGCATAAGGTCATAATTTAGCCAGAAT CATCAGCTTGT
[0300] > P. xylostella- Cactus (PxCac); Px016665 (SEQ ID NO: 122) CAATGCTGCGGAGGGTCTATGCGGGTGGACACCTCTACACGTAGCGGCGGCGCGAGGCGA
CGTCGACACGGCTCGCTACTTGCTCGAGAAGTGCGCTGGCGTCGATCCCTCTGCCCTGGA CTACGCCGGTCGTACGGCCAGGAAACTGGCGTTGAAGAATAAAGCGGCCGCCCTGTTTGA CGGCAGTGAGGGCAGCGAGGAGGAGGATAGTGACAGTGAGGATGAGATGCTTCTGGAAAG CGACCAGAGTCTGTTCGACCGGATCCGTGACGGTATGAACGCCATCAACGTCGCCTGA
[0301] > P. xylostella- Dorsal (PxDor); PxOO0110 (SEQ ID NO: 123) TAACCTGTGCCAGCAACCAATGGCTCCTATGGCGCAGCAGCTGATGGACCCGTCCCCCAG
CGACCCACCCTCCATCACGGGGCTGCTGATGGATCGCCCGGACCAGCCCTACTCCGGGGA GCTGTCTGGACTCTCCGCCCTGCTGGCTGAGGCAGCCCCCGCAGAGATGCTCAGCGATAG CCTCAACAGACTGTCTACGGGGGACTTGTTGAGACAAGTTGATATGTGA
[0302] > P. xylostella- Beta 1-3 glucan recognition protein 2 (PxPGRP2); Q8MU95.1 (SEQ ID NO: 124) GAAGGATTGAAGTGAGCGCCAGAATGCCGCGCGGTGATTGGTTGATTCCAGATATTCTGC
TGGAGCCGAAAGAAAACCTTTACGGAGTACGCAATTACGCGTCAGGTCTACTCAGCATAG CCTCAGTCAGAGGAAACACTGCTTACTCGAAGACCCTCAAAGGAGGCCCCATACTGTGTG ACAAGGAACCGCAGAGAAGTGCCAAGTTGAGCGAAAAAGTTGGATATGACCATTGGAATA
.0 AAGCCTTCCATAACTACACCATGATTTGGGCACCAAGTGGCATCACCATGCTGGTGGACG
GCGAGCAGTACGGGGACATCCGTCCCGGCGACGGCTTCAGCCAGGACCCGGCGGTGAGCA GCGTGGTGGCCGCGCCGCAGTGGCTGAAGGGCACCAGCATGGCGCCCTTTGATGTTATGT TCTACATATCCCTTGGTCTCCGCGTGGGCGGAGTGAACGACTTCCCCGACACTCCTGAGA AGCCGTGGAAGAACAAGGCCACTAAAGCCATGCTGAATTTCTGGAACGCCCGGGAACAGT GGCAGAGCAGCTGGTTTGAGGACACCACTGCACTCCTCATAGACTATGTCAGGGTTTATG CGCTGTGA
[0303] > P. xylostella- Chitin synthase 1 (PxCHS1); KX420688.1 (SEQ ID NO: 125) CGTATCTCGCACGACCTGAAAGAGCTGCGAAACTCATCCGTCTTTTCCTTCTTTATGATC
AATGCCCTCTTTGTTCTCATCGTATTCTTGCTGCAACTGAACAAGGACAACCTCCACATA AAGTGGCCCTTCGGAGTCAAAACTAACATTACGTATGATGAGGTGACGCAAGAGGTGCTG ATCTCCAAGGATACCTGCAACTAGAGCCTATTGGTCTGGTGTTCGTGTTCTTTTTCGCAT TGATTTTAGTCATCCAGTTCACTGCCATGTTGTTCCATCGATTCGGAACTTTGTCGCATA TATTATCGTCTACGGAACTGAACTGGTTCTGCAATAAGAAGGCGGAAGACTTATCTCAAG ACGCACTGCTAGATAAGAATGCGATAGCAATAGTGAAGGATCTCCAGAAACTAAACGGGC TCGATGACGGGTATGACAATGACTCGGGGTCGGGCCCGCACAATGTGGGAAGGAGAAAGA CGATACACAACCTGGAGAAAGCGAGACAGAAGAAGAGGAACATAGGAACGCTCGACGTCG CTTTCAAGAAGCGATTCTTCAACATGAACGCTAATGAAGGACCAGGAACACCAGTTCTGA ACCGCAAGATGACGTTGCGAAGAGAGACGTTGAAGGCGTTGGAAACGAGGAGGAATTCTG TGATGGCCGAACGAAGGAAGTCGCAAATGCAAACACTTGGAGCTAACAACGAATATGGAG TCACTGGAATCTTAAACAACAACCCAGCGGTGATGCCGCGCCACCGGCCGTCGACAGCCA ACATTTCGGTCAAGGACGTCTTCGCGGAACCCAACGGGGGACAAGTGAACCGAGGGTACG AGACCACGCACGGCGACGAGGGAGACGGCAACTCCATCAGACTGCAGCCGAGAACCAACC AGGTCTCCTTCCAGGGGAGATACCAATAA
[0304] > P. xylostella- Beta tubulin (PxPTUB); KX420688.1 (SEQ ID NO: 126) GAAGGAGGTCGACGAGCAAATGTTGAACATCCAGAACAAGAACAGCAGCTACTTCGTCGA
ATGGATCCCGAACAACGTCAAAACGGCCGTGTGCGACATACCGCCTCGTGGACTGAAGAT GTCTGCCACCTTTATCGGGAACACGACAGCAATCCAAGAGCTCTTCAAGAGGATTTCTGA GCAGTTCACTGCTATGTTCAGGAGGGAAGCGTTCCTCCACTGGTATACTGGTGAAGGCAT GGACGAGATGGAGTTCACAGAGGCGGAGAGCAACATGAACGACCTGGTCTCCGAGTACCA
.0 GCAGTACCAGGACGCCACGGCTGAAGACGAGGGAGAATTCGACGAGGATATTGAAGACGA
GTGA
[03051 > S. frugiperda-Peptidoglycan recognition protein 1 (SfPGRP1); repc7951 (SEQ ID NO: 127) CCTGATTGGTGGAAACGGGAGAGTTTATGAAGGAGCCGGCTGGCATCACGTTGGGGCCCA
TACTTTGGGATACAATGCAAGATCTGTGGGGATCTCCTTCATTGGCGATTTTAGAACAAA ATTACCAACACCCGAAGCACTGAAAGCCTTCAACAGTCTCCTGGAATGTGGAGTCACGAA CAATTATCTGTCAAAGGACTATCACCTGGTGGCCCATAGTCAGCTCTCTATGACTGACAG TCCYGGAGACATGYTGAGGAAGCAGGTGGAATCGTGGCCTCMTTGGCTGGATAATGCCAA AGACATACTTAAGTAGAAPAAGACTAAACGCCGTACTTTGAGCCATTTAATGGTTACTTA ACCCAGTCCTTAGCAATTTGATACAAGGCCAATGTCTCTAAGGGCGGCAGTAAAGGTCAA AACACATTTAATGAGTGTGTTTAAGATTTTGCTAGTGAAAATTGTTTTGAAGTACGTATT TGATGTAAGTGATGATATCAGTACCCTTAGTATGAGTTTGCTTTACGTTCCACGAGATGG AAACGAGAGCGCGTTCGGCGCTCTGATTGGTTCGTTCATTCATGCCGGCC
[03061 > S. frugiperda-Attacin (SfAtta); repc9395 (SEQ ID NO: 128) GCCAGACATCTACCACAGGACCACTCAACGTACGACCAAGTACAACTCCTCGGGTTCGAC
GAAGATGGACGACCAGTGTTTGAGCACGAAGACTTACTCCCAGAACTAGAGGAGTCCTAC CAGCCAGAGCACCTGGCGAGGACTCGCAGACAGGCGCAGGGCAGCGTCACCCTCAACTCC GACGGCGGCATAGGCCTGGGCGCTAAGATCCCGCTCGCACACAACGACAAGAATGTGGTG AGCGCCATCGGCTCCATGGACTTCAACAACAAGTTGCAGCCTGCTTCCAAGGGCTTCGGT CTGGCTCTGGACAACGTCAACGGGCACGGACTGACGGTGATGAAGGAAAGTATCCCCGGG TTCGGGGACAGGCTGTCGGGCGCTGGCAAGCTGAACGTGTTCCACAACGACAACCACAAC GTGGCCGTGACCGGCTCTCTCGCCAGGAACATGCCCAGCATCCCGAACGTGCCCAACTTC AACACGTACGGCGGGGGCGTCGACTACATGTACAAGAACAAGGTGGGAGCGTCTCTGGGC ATGGCCAGTACTCCGTTCTTGGACCGCAAGGACTACTCCGCGATGGGCAACCTGAACCTG TTCCGCAGCCCGACCACTACCGTGGACTTCAGCGGCGGCTTTAAGAAGTTCGAATCTCCC TTCATGAGCAGCGGCTGGAAGCCTAACTTCGGCCTTACTTTCGGCAGATCTTTCTAGATA TATTTTGTAATCTAAATTTAACTTTAACTTTGTTGTATAATATTTTGTCGAATTAAGATC AGTATTGTTCATACTAATATTATATTATCAGTGTTTCTTATAAATTAA
[0307] > S. frugiperda-Hemolymph proteinase 10 (SfHP10); c12881 (SEQ ID NO: 129) CAAGGCTCGTACCTTGCAGTTGAACCGACAATCTATTCCTAAAGCCTTTTTAAGGTCAGG
AAAAATAGTTCCTACATCTAAATGCAGTAGAATTTGCGAAACGAATTTAAATAAAAATGG CGTCGATTGTGTTTGTGATTTTGTGTGTTACCGTCGCTGCGGTGAAAAGCGCGATTTTAA ACCCGTGGAGTAAAGTTGAGGCCAACAAATGTGGTGTAGAAGCCAGTACTAACTTGGTCC ATCACAATCCATGGTTGGTCTACATCGAGTATTGGCGTGGAAACTCAGATACTGAGATCC GATGCGCCGGTACTTTAATCGACAGCAAACATGTCGTCACAGCTGCCCACTGCGTTAGGA CTCTGAAGTTTAGTCATTTGATCGCCCGTCTTGGCGAATACGACGTAAATTCTAAGGAGG ACTGCGTTCAGGGCGTGTGTGCCGATCCCATCGTCAGAATCAAGGTGGCTGAGATCATCG TGCATCCTAACTACAGCAACCGGGAACA
[0308] > S. frugiperda-Trypsin like serine protease (SfTSP); repc48453 (SEQ ID NO: 130) AGCAACAAAATGCGTGTCCTCGCTTGCTTGGCCCTTCTCTTAGCTGTGGTAGCAGCCGTC
CCCTCCAATCCCCAGAGGATTGTGGGTGGTTCGGTCACCACCATTGACCGGTACCCCACC ATTGCATCCCTGCTGTACTCGTGGAACTTGAGTTCCTACTGGCAGGCGTGCGGTGGTTCC ATCTTGAACAACCGTGCCATCCTTACTGCTGCCCACTGCACAGTTGGTGACGCCGCCAAC AGATGGAGAATCCGTCTTGGCTCCACCTGGGCCAACAGCGGTGGTGTCGTTCACAACGTC AACACTAACATCGTCCACCCCTCATACAACTCTGCAACTTTGAACAACGACATCGCTATC CTCCGCTCCGCCACCACCTTCTCCTTCAACAACAATGTTCAGGCTGCCTCCATTGCTGGT GCCAACTACTTGCCCGGTGACAACACCGCCGCCTGGGCCGCTGGATGGGGAACTACCTCC GCTGGTGGCTCTAGCTCTGAGCAGCTCCATCACGTTGAGCTGCGCATCATCAACCAGGCT ACTTGCAAAAACAATTACGCTACCCGCGGTATCACCATCACCTACAACATGTTGTGCTCT GGCTGGCCCACCGGTGGTCGCGACCAGTGCCAGGGTGACTCTGGTGGTCCTCTCTACCAC AACGGCATCGTTGTTGGTGTCTGCTCTTTCGGTATTGGCTGTGCTCAG
[0309] >S. frugiperda- C Type Lectin 6 (SfCTL6); Joint2_ repc448 (SEQ ID NO: 131) AACAGTTTTCTATTGGCAGTCAAAGACTTCAGTCGAAAAATAATCCTCATCAGAGTCGTG
AAGCAAGGTGCCCAAAATATAATGTAACCTAGATACCTATTAATAAATTATTTGTCAACC
.0 AAAACGTTACGTTCAAAGTCCTTAAAATCAAAATATCTTATGATTAGTTTTGATTTAAAA
ATAGAGGTTCGAAATCGCCAACCCAAATAGGTTTAGTTTACGATTCAGGAAAAATCCTAA CGTAGGGAAACATTATTTTACAAGACTTTTGGCTTAAAAACTTTGAGAACCAATGTCAAA TTTGATAATAACTAATGAGGTATAAAAGCTTGATCCTATTAGGACTTATTTTCATAACAC CATCGAGTTTGTATTTAATRRAGACGTGKGTTAACTAACAACATGAAGACCGGTGTAAAA TATTCTGTTMTTTGGATATTCTCTCTATTYTGCTATATAGAGGCAACATTTCGTTGTGAC TACACGTACAGCAAGGAAGCGAAGGGCTGGTTCAAACATGTGGTGATACCAGCTACTTGG GCTGACGCACGWCTGCACTGCACGTTGGAAGGTGCAACGCTGGCTTCTCCACTCAACCAG GCTATMAGTAATGAGATGCAGTCCMTCCTGGCPAACCTCTCGGCGCTGCAATCAGAAGTC TTCACTGGAATTCACRCGACTKTTTCACGRMRCAACTTATATCATACYATYGAAGGTATA CCTCTTAGTAAAATTCCATTAGATTGGGCAACAAATGAGCCAAATGGTGGGAGAGATGAA AACTGTATCACGTTTAACTCCGATGGCCAAGCGGCAGACAGATCCTGTAARGAGACTCGA CCTTACATCTGCTACCGACACACWACTAAAGTGACTGTGKCCAATGAATGTGGGACTGTA GATCCTGAATACAATTTGGATAAAAGAACGGGCKCYTGCTATAAGTTCCACACRGTACCT CGCACGTTCGAGCGTGCCAACTTCGCGTGTTCTGCTGAAGGTG
[0310] > S. frugiperda- Cecropin (SfCec); repc42380 (SEQ ID NO: 132) TTCGTGTCGTATCACTAGAGTTCGAAATACAAAATAATAATACATTTATTATTTTGCCAT
AATTAATAATAAAGTTATTTTATTTCATAATAATAATGAATTTCACAAAGATATTTTGTT TGTTTTTGTCTTGCTTTGTTTTGATGGCGACCGTGTCAGGAGCTCCTGAACCGAGGTGGA AATTCTTCAAGAAAGTGGAGAAGTTGGGCCAAAACATCCGCKATGGTATCATAAAGGCAG GACCCGCAGTGGCCGTGGTGGGATCAGCRGCAGCCATWGGAAAGTGAKCCCTACGACCTG AGACATGAAGACTAATATCCAYTAAAATAASAATATTGAGGCKTATAATATTAATTTATT RTRTTTGTAAATTAAATTATTTGTAAGATAA
[0311] > S. frugiperda- Beta 1, 3 glucanase recognition protein (SfPGRP2); EF641300 (SEQ ID NO: 133) GCAACAATCGCACCAACTCTTTCGTGCGCAGTGGAAGTTTGTTCATCCGTCCCTCTCTAA
CATCAGACGAGTTCGGAGAAGCTTTCCTCTCATCTGGACACTGGAACGTCGAGGGTGGTG CTCCTGCTGATAGATGCACAAATCCACAATGGTGGGGTTGCGAGAGAACAGGCACGCCGA CCAACATTTTGAACCCAATCAAGAGTGCTCGTGTCCGTACCGTCAATTCCTTCAGCTTCC GTTACGGACGCCTCGAAGTCCGCGCTAAAATGCCCGCCGGAGATTGGATTTGGCCAGCTA T CTGGTTGATGCCTGCGTACAACACTTACGGTACTTGGCCCGCATCAGGAGAGATTGACT TAGTTGAGTCCCGAGGCAACCGTAACATGTTCCACAATGGTGTCCATATCGGTACACAGG AAGCAGGCTCGACCTTGCACTACGGACCTTACCCAGCGATGAACGGTTGGGAGCGCGCCC ATTGGGTCAGAAGGAACCCT
[0312] > S. frugiperda- joint2_repc16438 (Sfrc16438); Un-annotated (SEQ ID NO: 134) TCGTTCTCCTCGTCGCTTTCTTGGGGACCTCATGGTTTACGGGAGATGTTTCTGCGAGTC
CGCGGCCGCAAGAGCCGCGTGTGGATCAAAATCCGAATCAGGTGTCACCTTATGGAGGGT CCGGGTACCACGCACCTCCGCAGTACCAACCGCAGTACCAACCACAGCCGTACTACCCAC AGCCGCAGTACTACCCACAGCCGTACTACCCACCTCCTCAGTATTACCCACCGCAGCCAC AAACACCTGAGAATGCTCCACTCATAAACACATGGAATGGTTTCCACGACTGGGCTCAGA ATATCGTTCAAAGTGCTTTGGGGCAGAAATTCCCGAAAGGTAGACAGTAACTTTTTAATT GTCAATTGAAGATAAGGCCCATTTCACCAACTGCTGTTTAATTTTAAGGAGCTCCTAAAC TAACATAGGTGACACTTAGCGATATTCTGGATTTTTTGTGAACGTATAAATAATATCCAA ATGTAAAAGATAAGAGGCCAAGAA
[0313] > S. frugiperda- Chitin synthase B (SfChsB);AY52599) (SEQ ID NO: 135) GAATTTAGGAGCAGCGTGCGGGCGCATCCATCCTGTGGGCTCAGGCTTCATGGCATGGTA
TCAAATGTTCGAGTACGCTATTGGTCATTGGCTGCAAAAGGCGACTGAACACATGATTGG CTGTGTACTCTGTAGCCCTGGATGCTTCTCCCTCTTCAGAGGAAAGGCTTTGATGGACGA CAACGTTATGAAGAAATATACCTTAACTTCCCACGAGGCACGACACTATGTGCAATACGA TCAAGGCGAGGACCGTTGGTGCACGCTACTGCTGCAGCGCGGGTACCGCGTGGAGTACAG CGCGGTGTCGGACGCGTACACGCACTGCCCCGAGCACTTCGACGAGTTCTTCAACCAGCG CCGCCGCTGGGTGCCCTCCACGCTCGCCAACATCTTCGACCTGCTCGGCAGCGCCAAGCT CACCGTCAAGTCCAACGACAACATCTCCACCCTCTATATAGTCTATCAGTTCATGTTGAT AGTGGGTACGGTGTTGGGTCCCGGCACGATCTTCCTGATGATGGGGGGAGCCATGAACGC CATCATTCAGATCAGCAACGCGTACGCGATGATGTTGAACCTCGTACCACTCGTCATCTT CCTTATAGTCTGTATGACTTGTCAGTCAAAGACGCAGCTCTTCCTCGCTAACCTCATAAC ATGCGCATACGCAATGGTGATGATGATCGTGATAGTGGGGATAGTTCTGCAGATAGTGGA GGATGGATGGCTGGCTCCGTCCAGTATGTTCACAGCTTTAATATTCGGTACATTCTTCGT CACCGCGGCACTACACCCGCAAGAGATCAAATGTTTGTTGTTCATAGCAGTGTACTATGT
.0 AACCATCCCTAGTATGTACATGTTGTTGATCATATACTCCATCTGTAATCTCAACAACGT
ATCCTGGGGTACCAGGGAGACACCGCAGAAGAAAACTGCTAAGGAAATG
[0314] > T. castaneum- Peptidoglycan recognition protein 2 (TcPGRP2); XM_965754.3 (SEQ ID NO:136) A TGG CAVG C3T GACCC T T TAACAAA TACCCAACA T CCGA T CAAGA T TA T TACA T CCA
CA T AAGAAiAGT T TGCGAG T T CAGGA TAAAATT C T T TA T TCTAG T TI TTA T TI' TTCAA T T
CTAATTACCGG5ACTAGCCATTGGCCTCTATCTCCTTCAACTGAGGACACGAATGGAAA 3ACA T GCGk T T GACGT"T "CCG TAGGG T UUT' T TA T T TTATCCI TA CAA TA"CCAC -- T GCTGCAGGAGTCTATAATATTACAGTTCGGGAACAGTGGAAGCTCACGTCCCTTCATCA
AGCTGCGGC.AGCAAATCC:CACTGTGCcCAAAGCCCCAGGAACTACAATTACCATATG3 CTGCAGTGGAAAGAACCT GACA TCT CC TACAAT TTCA TAAT GAC TCAGAT GGCAGAATT TTCGAGGGGAGAGGA T GGGAC:TT T GAAACT T T TT T T CAAAT GTACGG T TAATGATAC T
GTGACAGTTGC.TTCTGACGAATTT GCGAAAGCACCGACGTTTAGACAAGCTGAA CGGCAAAAAT GT T CCT GGAAG T
[0315] > T. castaneum- Beta 1-3 glucan binding protein (TcpGRP2); XM_966587.4 (SEQ ID NO:137) AC AACGTCCCGACCCTCAATCCAAGCATTCACCCCCTGCTTCAAAGTGAG
CATCCCTCATACTGAACCACCAATTATTCGCCTTTCATGGAAATATTAATAAACCCCT GCACGGTCTCCAGCCGGACATTTTCCCAAGCCGCCTCCAAAGGGAGGGAGACGAGTG
GGTGTTCcCAAGACTCCAGTCCA TTAAACCCGGGAGATAPAATCTATTATTGGCTCTT TATCATTAAAAGACCCT GCTACAGATACGACATCACGCCGACTACGAAGCC TAAAGCT TT AGCA CCG ACG CACG"CCCCCCAA
TCTCGGCATTTGCGAAkAGGTGATGGTAkATCTCACGCGAAGCTCTCGATTTGCAGCA
AGAAA T T GAG T CC"C T TAGGGATATGAAGACTAT TT TGGAGTAC G T CAGAAGCACAC TGATACCGCCACTACCTCCACTTCGACGCC TTAT A'TAAGAT GAGACAACTT TCC T TAAGCAA CT T TAT TAAAGAAAACT TACAAAAAGCAGGA 'TCAAAA TGT GACGAGGCAGCAGAA T GGAA T GG T CAAG T T T CAAG T CGAG T T T GAGAAAAAG T T GGA CGGT T TAAAGCGGCCAAAAGAAAACTC'_AA G TCGTCGAGC-T T TAC GAT CACGSTAT TA/A
[0316] > T. castaneum- midgut protein (TcMDGP); XM_971351 (SEQ ID NO: 138) CCTGAAGAGA TGCCCAGCTCGAAGAAAT CAG TA T CAGCTCGATCTCCTAGCCAAATCGAAA
GAAGT TTTAC TAGANAT TAGCCCGAACTTTCGATGAAAATCCTGAAACCGGCCTCTCAGGAAC GAGGACAGCT GGC T GCAGAGAG TGCAGAAAAT GGACGAT TTAT T CAT CAGCT GGACGAC
CTC TTATCCAGACGATACCAkAATCGACTCACGCAACCCCAAATTC CCCTGGAC
C CCGACTTCCGCATCGTGGAC'ITTCA.GCGACCITCCCATT CCTCGACCTAAAC CTAAACCCTCGGGCAATCTC AATCGAGGCACGTACGAAGCCAACAACACCACACT CACG CGATGGCTCCCGGTATCTCACATTGGTCGAATCGTGATCGGTTTTCAAACGTCCGACG AACGAAAGCGGACC GTGCT"'CGAGCAGGATTCCTCCCAGATTATGAAATT AGATATGAGCCGACGGACCCTGTTATAGGGTTAGCTATCACCCGGATGGCGAAATGAG CCTGCAAAATGACATTAGCAATT CAGATAT TGAGGCCACCTAGGCAAGACCCCCCTGG TG CAG T T GACT GAGATAT T G T CCACC T G T T GCATAGCAAT T GGCCGAGG T T G TAGT GGA T
[0317] > T. castaneum- Chitin synthase 2 (TcCHS2); EFA 10719.1 (SEQ ID NO: 139) ATGGCGGCGCGTCATCGCTTTGCCACAGGGAGCCCTGAGGAAACAGAGCCCCTGTATTCG
TCGACGCAAATGCCCGAAAAAGTCCGGGAAAAATGGAACGTCTTCGACGACCCCCCAAGA GAGCCCACTTCGGTTCCGAAGTCAAAAGAACCTACATCGAGTGGGGGGTGAAGTTTTTGA AAGTTGTGACAATCATAACTGTGTTTTTTGTGGTCCTTGGTGCTGCAGTGGTTTCGAAAG GGACAACCTTGTTCATGACGTCACAAATAAAAAAGAATGTGACAAGGGCTTATTGCAACA AAAAGATAGACCGCAACCTCCAATTCGTCGTCTCCCTCCCCGAAGTGGAGCGCGTGCAAT GGATCTGGCTCCTCATTTTCGCTTACTTGATCCCCGAAGTGGGTACCTGGATCCGCGCCG TCCGCAAATGCCTCTACAAGCTCTGGAAAATGCCCTCCCTCTCCGAATTCCTCTCCCTCT TCGGCACGGAAACGTGCCCCGCCATCGGAAGCGCAATTTTGATATTCGTCGTCCTCCCCG AGCTGGACGTCGTCAAAGGGGCGATGCTCACAAACGCGGTTTGTTTCGTGCCCGGAGTTG TGGCAATGTTCTCGCGCAAACCGTGCTCCATAAACGAGAACCTGAAAATGGGGCTGGACA TCGCCAGCATAACTGCACAGGCGTCAAGCTTCGTGGTGTGGCCCTTGGTTGAAAATAACC CGACCTTGTACCTAATCCCCGTTTCCGTGATTTTGATTTCGGTGGGT
[0318] The breadth and scope of the present disclosure should not be limited by any of the above-described exemplary embodiments, but should be defined only in accordance with the following claims and their equivalents.
References
1. http World Wide Web internet site "fao.org/3/a-avOl3e.pdf'.
2. Polaszek A. (1998). African Cereal Stem Borers: Economic Importance, Taxonomy Natural Enemies and Control. Wallingford, UK: CABI. 530 pp.
3. http World Wide Web internet site "cnbc.com/2015/05/08/insects-feast-on-plants endangering-crops-and-costing-billions.html".
4. Khan ZR. et al. (2014). Achieving food security for one million sub-Saharan African poor through push-pull innovation by 2020. PhilosophicalTransactionsof the Royal Society B: BiologicalSciences, 369 (1639), 20120284.
5. Tabashnik BE. et al. (2013). Insect resistance to Bt crops: lessons from the first billion acres. Nat. Biotech. 31, 510-521.
6. Campagne et al. (2013). Dominant inheritance of field-evolved resistance to Bt corn in Busseola fusca. PLoS ONE 8(7):e69675. .0 7. http World Wide Web internet site "dtnpf com/agriculture/web/ag/news/article/2016/08/10/rootworm-resistance- pyramided bt".
.5 8. Terenius 0. et al. (2011). RNA interference in Lepidoptera: An overview of successful and unsuccessful studies and implications for experimental design. Journal of Insect Physiology 57, 231-245.
9. Gayatri Priya N. et al. (2012). Host Plant Induced Variation in Gut Bacteria of Helicoverpaarmigera. PLoS ONE. 7(1):e30768.
10. Penuelas J and Terradas J (2014). The foliar microbiome. Trends Plant Sci. Volume 19, Issue 5, 278 - 280.
11. Casanova-Torres and Goodrich-Blair (2013). Immune Signaling and Antimicrobial Peptide Expression in Lepidoptera. Insects. 4, 320-338.
12. Tang X. et al. (2012). Complexity and Variability of Gut Commensal Microbiota in Polyphagous Lepidopteran Larvae. PLoS ONE. 7(7):e36978.
13. Ryu J1. et al. (2008). Innate immune homeostasis by the homeobox gene caudal and commensal-gut mutualism in Drosophila. Science. 319, 777-82.
14. Shrestha S. et al. (2009). An inhibitor of NF-kB encoded in Cotesiaplutella bracovirus inhibits expression of antimicrobial peptides and enhances pathogenicity of Bacillus thuringiensis. JournalofAsia-PacificEntomology. 12, 277-283.
15. Buchon N. et al. (2013). Gut homeostasis in a microbial world: insights from Drosophila melanogaster. Nat. Rev. Microbiol. 11, 615-626.
16. Karlin S and Altschul SF (1990). Method for assessing the statistical significance of molecular sequence features by using general scoring schemes. Proc. Natl. Acad. Sci. USA. 87,2264-2268.
17. Watson JM. et al. (2005) RNA silencing platforms in plants. FEBS Letters. 579, 5982 8987.
18. Zhong X. et al. (2012). A Toll-Sptzle Pathway in the Tobacco Hornworm, Manduca sexta. 42(7), 514-524.
19. Zhang X. et al. (2015). Phylogenetic analysis and expression profiling of the pattern recognition receptors: Insights into molecular recognition of invading pathogens in Manduca sexta. Insect Biochem. Mol. Biol. 62, 3 8-50.
20. Cao X. et al. (2015). The immune signaling pathways of Manduca sexta. Insect Biochem. Mol. Biol. 62, 64-74. .0 21. Wang et al. (2006). Interaction of 0-1,3-Glucan with Its Recognition Protein Activates Femolymph Proteinase 14, an Initiation Enzyme of the Prophenoloxidase Activation System in Manduca sexta J. Biol. Chem. 281(14), 9271-9278.
.5 22. Ward N and Moreno-Hagelsieb G (2014). Quickly Finding Orthologs as Reciprocal Best Hits with BLAT, LAST, and UBLAST: How Much Do We Miss? PLoS ONE 9(7), e101850.
23. Timmons L. et al. (2001). Ingestion of bacterially expressed ds RNAs can produce specific and potent genetic interference in Caenorhabditiselegans. Gene 263, 103-112.
24. Kanost MR. et al. (2016). Multifaceted Biological Insights From a Draft Genome Sequence of the Tobacco Hornworm Moth, Manduca Sexta. Insect Biochem. Mol. Biol. 76, 118-147.
25. Odman-Naresh J. et al. (2013). A lepidopteran- specific gene family encoding valine-rich midgut proteins. PLoS ONE 8:e82015.
26. Engel P and Moran NA (2013). The gut microbiota of insects - diversity in structure and function. FEMS microbiology reviews. 37(5), 699-735.
27. Siepel A. et al. (2005). Evolutionarily conserved elements in vertebrate, insect, worm, and yeast genomesGeinoneRes. 15,1034-1050.
28. Pauchet Y. et al. (2010). Pyrosequencing theManduca sexta larval midgut transcriptome: messages for digestion, detoxification and defence. InsectMolBiol. 19, 61-75.
29. Kim SH and Lee WJ (2014). Role of DUOX in gut inflammation: lessons from Drosophila model of gut-microbiota interactions. Front. Cell. Infect. Microbiol. 3, 116.
30. Lee WJ and Hase K (2014). Gut microbiota-generated metabolites in animal health and disease. Nat. Chem. Biol. 10, 416-424.
31. Brummett et al. (2017). The immune properties of Manduca sexta transferrin. Insect Biochem Mol Biol. 81, 1-9.
32. Xia Xu et al. (2012). Manduca sexta Gloverin Binds Microbial Components and is Active against Bacteria and Fungi. Dev Comp Immunol. 38(2), 275-284.
33. Gunaratna RT and Jiang H (2013). A comprehensive analysis of the Manduca sexta imnmunotranscriptome. Dev, Comp. mmunol 39, 388-398,
34, Broderick NA. et al. (2009). Contributions of gut bacteria to Bacius thuringiensis induced mortality vary across a range of Lepidoptera. BMC Biol. 7:11.
35. Gregory R. Richards (2008). Xenorhabdus nematophila lrhA is necessary for motility, .0 lipase activity, toxin expression, and virulence in Mnduca sexta insects. Journal of Bacteriology. 190, 4870-4879.
36. Timmons and Fire (1998). A. Specific interference by ingested dsRNA. Nature. 395, 854.
.5 37. Kamath RS. et al. (2000). Effectiveness of specific RNA-mediated interference through ingested double-stranded RNA in Caenorhabditiselegans. Genome Biol. 2, 1-10.
38. Cao et al. (2018). A systematic study of RNAi effects and dsRNA stability in Tribolium castaneum and Acyrthosiphon pisum, following injection and ingestion of analogous dsRNAs. Int. J. Mol. Sci.19, 1079.
39. Rajkumar AP et al. (2015). Experimental validation of methods for differential gene expression analysis and sample pooling in RNA-seq. BM/CGenonics. 16(1), 548.
40. Zhang et al (2015). Full crop protection from an insect pest by expression of long double-stranded RNAs in plastids. Science. 347, 991.
41. Kim SR, Yao R, Han Q, Christensen BM and Li J (2005). Identification and molecular characterization of a prophenoloxidase involved in Aedes aegypti chorion melanization. Insect molecular biology. 2005;14(2):185-194.
42. Packey CD and Sartor RB (2009). Commensal Bacteria, Traditional and Opportunistic Pathogens, Dysbiosis and Bacterial Killing in Inflammatory Bowel Diseases. Current opinion in infectious diseases. 2009;22(3):292-301.
43. Arakane et al. (2005). The Tribolium chitin synthase genes TcCHS1 and TcCHS2 are specialized for synthesis of epidermal cuticle and midgut peritrophic matrix. Insect MolecularBiology 14(5), 453-463.
44. Morris et al. (2009). Tribolium castaneum larval gut transcriptome and proteome: A resource for the study of the coleopteran gut J. Proteome Res. 8(8):3889-98.

Claims (20)

CLAIMS What is claimed is:
1. A method of silencing an insect immune response gene, an insect gene encoding structural components of an insect midgut, or both, the method comprising providing for ingestion: (a) an isolated double stranded RNA (dsRNA) molecule, or a dsRNA molecule in a host cell, in a transgenic or transplastomic plant or cell, organelle, or part thereof, in a microbial conduit, or in an insecticidal composition, wherein the dsRNA molecule comprises a nucleic acid sequence complementary to about 21 to 2000 contiguous nucleotides of a target gene sequence comprising a nucleic acid sequence of SEQ ID NO: 76, wherein the dsRNA molecule silences the target gene when ingested by an insect; (b) an siRNA molecule derived from the processing of the dsRNA molecule; (c) a polynucleotide, a construct, or a dsRNA encoding segment encoding the dsRNA molecule; or (d) a combination of (a)-(d).
2. A method of protecting a plant from an insect pest of the plant, the method comprising topically applying to the plant (a) an isolated double stranded RNA (dsRNA) molecule, or a dsRNA in a host cell, in a transgenic or transplastomic plant or cell, organelle, or part thereof, in a microbial conduit, or in an insecticidal composition, and providing the plant in the diet of the insect pest, wherein the dsRNA molecule comprises a nucleic acid sequence complementary to about 21 to 2000 contiguous nucleotides of a target gene sequence comprising a nucleic acid sequence of SEQ ID NO: 76, and wherein the double stranded RNA molecule silences the target gene when ingested by an insect; (b) an siRNA molecule derived from the processing of the dsRNA molecule; (c) a polynucleotide, a construct, or a dsRNA encoding segment encoding the dsRNA molecule; or (d) a combination of (a)-(d).
- 142 89015902.1
3. The method of any one of claims 1-2,wherein the microbial conduit comprises plant growth promoting organisms, normal commensal and/or symbiotic microorganisms associated with the target insect pest or parasites, and/or natural enemies of the target pest or pest target host or host cultivation range etc. from an insect or parasite, and/or natural enemies of the target pest engineered or identified from natural populations containing microbial conduit to produce and/or deliver dsRNA and/or drive the transmission of such microbial conduits into natural populations of insect pests as a control option.
4. The method of any one of claims 1-3, wherein the dsRNA molecule is bound to a synthetic carrier.
5. The method of claim 4, wherein the synthetic carrier comprises chitosan, liposomes, carbon quantum dots, biodegradable particles of plant, or soil.
6. A method of producing a transgenic or transplastomic plant, the method comprising: (a) transforming the plant with a polynucleotide encoding a dsRNA, a construct or a dsRNA encoding segment encoding the dsRNA molecule, or both; (b) regenerating a plant from the transformed plant cell and/or organelle; and c) growing the transformed plant under conditions suitable for the expression of said double stranded RNA molecule; wherein the transformed plant of (c) is resistant to a plant pest insect compared to an untransformed plant and wherein the dsRNA molecule comprises a nucleic acid sequence complementary to about 21 to 2000 contiguous nucleotides of a target gene sequence comprising a nucleic acid sequence of SEQ ID NO: 76.
7. The method of any one of claims 1-6, wherein ingestion of the dsRNA, the siRNA molecule, the polynucleotide, construct, or dsRNA encoding segment encoding the dsRNA molecule, or any combination thereof silences the target gene to thereby induce (a) a melanotic response; (b) results in perturbation of gut microbial homeostasis; (c) results in defective clearance of opportunistic microbes; (d) results in defective containment of gut microbes, or any combination of (a) to (d). - 143 89015902.1
8. The method of any one of claims 1-7, wherein the host cell is a bacterial cell, a yeast cell, or a fungal cell.
9. The method of any one of claims 1-8, wherein the target gene sequence includes at least one of the protein coding region, the 5' untranslated region (UTR), the 3' UTR, or any combination thereof.
10. The method of any one of claims 1-9, wherein the dsRNA molecule comprises a single RNA strand comprising an inversely repeated sequence with a spacer in between and where the single RNA strand can anneal to itself to form a hairpin loop structure; or wherein the dsRNA molecule comprises two separate complementary RNA stands annealed together.
11. The method of any one of claims 1-10, wherein the target gene is selected from the group consisting of Manduca sexta-Peptidoglycan recognition protein 2 (MsPGRP2), Plutella xylostella PGRP2, and Tribolium castaneum PGRP2.
12. The method of any one of claims 1-11, wherein the dsRNA comprises a nucleic acid sequence of SEQ ID NO: 76, or a fragment of at least about 21 nucleotides thereof; optionally.
13. The method of any one of claims 1-12, wherein the dsRNA molecule causes impeded growth, developmental progression, and/or mortality and the like of DBM, optionally wherein the DBM is a Bt resistant strain.
14. The method of any one of claims 1-13, wherein the construct encoding the dsRNA comprises a gene silencing sequence operably linked to one or more promoters for the expression of a dsRNA molecule that silences the target gene when ingested by an insect, optionally wherein the recombinant DNA construct further comprises an additional transcription regulatory region or an additional transcriptional regulatory element.
- 144 89015902.1
15.The method of claim 14, wherein the silencing of the target gene results in reduced appetite and/or developmental defects resulting in incomplete development and/or mortality and/or decreased reproductive success of the insect, optionally wherein the reduced appetite and/or developmental defects and/or mortality and/or reduced reproductive fitness of the insect is observed after sustained feeding for at least 24 hours.
16. The method of any one of claims 1-15, wherein the construct is an expression vector, and the expression vector can target single or multiple insect RNAi target genes or chimeric RNAi target genes.
17.A method of improving crop yield, the method comprising growing a population of transgenic or transplastomic plants comprising a polynucleotide encoding a dsRNA, a construct or a dsRNA encoding segment encoding the dsRNA molecule, wherein the dsRNA molecule comprises a nucleic acid sequence complementary to about 21 to 2000 contiguous nucleotides of a target gene sequence comprising a nucleic acid sequence of SEQ ID NO: 76 and wherein the population of transformed plants produces higher yields in the presence of pest insect infestation than a control population of untransformed plants.
18. The method of any one of claims 1-17, wherein the insect is of the order Lepidoptera, Coleoptera, Hemiptera, Blattodea, or Diptera.
19. The method of any one of claims 1-18, wherein the insect is Manduca sexta (M. sexta) (tobacco hornworm), Spodoptera frugiperda (fall armyworm), Ostrinia nubilalis (European corn borer), Plutella xylostella (Diamondback moth), Leptinotarsa decemlineata Say (Colorado potato beetle), Diabrotica spp. (Corn rootworm complex), Tribolium castaneum (Red flour beetle), Popilliajaponica (Japanese beetle), Agrilus planipennis (Emerald ash borer), Diaphorina citri (Asian citrus psyllid), Cimexlectularius (Bed bug), a cockroach or termite, or insect pests such as mosquitoes and flies.
- 145 89015902.1
20.The method of any of of claims 1-19, wherein the plant is selected from the group consisting of Zea mays L (corn), Sorghum bicolor (sorghum), Setaria italica (fox tail millet), Pennisetum glaucum (Pearl millet), Solanum tuberosum (potato), Oryza sativa (rice), Lycopersicon esculentum (tomato), Solanum melongena (eggplant), cultivars of the Brassica oleracea family, Citrus sinensis (Orange), trees of the Oleaceae family, and crops of Rosaceae.
- 146 89015902.1
This data, for application number 2018261075, is current as of 2023-04-12 21:00 AEST
AU2023202271A 2017-05-01 2023-04-13 RNAi approach for crop pest protection Pending AU2023202271A1 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU2023202271A AU2023202271A1 (en) 2017-05-01 2023-04-13 RNAi approach for crop pest protection

Applications Claiming Priority (5)

Application Number Priority Date Filing Date Title
US201762492556P 2017-05-01 2017-05-01
US62/492,556 2017-05-01
PCT/US2018/030506 WO2018204398A1 (en) 2017-05-01 2018-05-01 Rnai approach for crop pest protection
AU2018261075A AU2018261075A1 (en) 2017-05-01 2018-05-01 RNAi approach for crop pest protection
AU2023202271A AU2023202271A1 (en) 2017-05-01 2023-04-13 RNAi approach for crop pest protection

Related Parent Applications (1)

Application Number Title Priority Date Filing Date
AU2018261075A Division AU2018261075A1 (en) 2017-05-01 2018-05-01 RNAi approach for crop pest protection

Publications (1)

Publication Number Publication Date
AU2023202271A1 true AU2023202271A1 (en) 2023-05-11

Family

ID=64016305

Family Applications (2)

Application Number Title Priority Date Filing Date
AU2018261075A Abandoned AU2018261075A1 (en) 2017-05-01 2018-05-01 RNAi approach for crop pest protection
AU2023202271A Pending AU2023202271A1 (en) 2017-05-01 2023-04-13 RNAi approach for crop pest protection

Family Applications Before (1)

Application Number Title Priority Date Filing Date
AU2018261075A Abandoned AU2018261075A1 (en) 2017-05-01 2018-05-01 RNAi approach for crop pest protection

Country Status (12)

Country Link
US (2) US20200181639A1 (en)
EP (1) EP3618634A4 (en)
CN (1) CN110914429A (en)
AU (2) AU2018261075A1 (en)
BR (1) BR112019022872A2 (en)
CA (1) CA3061721A1 (en)
CL (2) CL2019003139A1 (en)
MX (1) MX2019012986A (en)
PE (1) PE20200691A1 (en)
RU (1) RU2019138709A (en)
SG (1) SG11201910115XA (en)
WO (1) WO2018204398A1 (en)

Families Citing this family (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2020176224A1 (en) * 2019-02-27 2020-09-03 Donald Danforth Plant Science Center Antimicrobial ncr2 peptides
CN109880826A (en) * 2019-03-20 2019-06-14 新疆农业科学院植物保护研究所 Application and product of the dsRNA in colorado potato bug prevents and treats
US20210321616A1 (en) * 2020-04-17 2021-10-21 Evolutta Agro Biotecnologia Ltda Process of selection of a nucleotide sequence for use as a phytosanitary product, plasmid or cosmid, host cell, process for the production of an rna molecule, rna molecule, use of an rna molecule, stabilized compound, phytosanitary product and method to eliminate or reduce the infestation of an insect, disease or weed in agriculture
CN111518803B (en) * 2020-04-27 2023-05-12 广西壮族自治区林业科学研究院 RNAi fragment and application thereof in regulation and control of lignin synthesis
CN116157019A (en) * 2020-05-13 2023-05-23 绿光生物科学公司 RNA-based control of lepidopteran pests
CN114854774A (en) * 2021-02-04 2022-08-05 南京吉星生物技术开发有限公司 Chordoporus punctatus chitin synthase 1(CHS1) gene and application thereof in aspect of RNAi mediated pest control
CN113430201B (en) * 2021-08-17 2022-04-01 中国林业科学研究院森林生态环境与保护研究所 Hyphantria cunea Iap gene dsRNA, bacterial expression bacterial liquid thereof and application thereof
WO2023034849A2 (en) 2021-08-31 2023-03-09 nanoSUR LLC HIGH MOLECULAR WEIGHT MODIFIED dsRNA COMPOSITIONS
CN113845579B (en) * 2021-11-15 2023-06-23 扬州大学 Preparation method of immune lectin for improving killing efficiency of beauveria bassiana on cotton bollworms

Family Cites Families (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6326193B1 (en) * 1999-11-05 2001-12-04 Cambria Biosciences, Llc Insect control agent
WO2005049841A1 (en) * 2003-11-17 2005-06-02 Commonwealth Scientific And Industrial Research Organisation Insect resistance using inhibition of gene expression
US20060021087A1 (en) * 2004-04-09 2006-01-26 Baum James A Compositions and methods for control of insect infestations in plants
PL2431473T3 (en) * 2005-09-16 2017-05-31 Monsanto Technology Llc Methods for genetic control of insect infestations in plants and compositions thereof
CA2630932A1 (en) * 2005-11-21 2007-05-24 Johnson & Johnson Research Pty Limited Multitargeting interfering rnas and methods of their use and design
EP2377939A3 (en) * 2006-01-12 2012-01-18 deVGen N.V. Transgenic plant-based methods for plant pests using RNAi
CN103484468B (en) * 2013-09-11 2015-04-08 华南农业大学 Diamondback moth peptidoglycan recognition protein, preparation method and application thereof
US10093945B2 (en) * 2015-11-13 2018-10-09 The United States Of America, As Represented By The Secretary Of Agriculture Double strand RNA-mediated RNA interference through feeding detrimental to larval Lymantria dispar (gypsy moth)

Also Published As

Publication number Publication date
BR112019022872A2 (en) 2020-05-19
CL2022002164A1 (en) 2023-03-10
CA3061721A1 (en) 2018-11-08
US20200181639A1 (en) 2020-06-11
MX2019012986A (en) 2020-08-06
CN110914429A (en) 2020-03-24
AU2018261075A1 (en) 2019-12-12
CL2019003139A1 (en) 2020-03-20
EP3618634A1 (en) 2020-03-11
US20240041914A1 (en) 2024-02-08
RU2019138709A (en) 2021-06-02
WO2018204398A1 (en) 2018-11-08
PE20200691A1 (en) 2020-06-11
RU2019138709A3 (en) 2021-09-22
EP3618634A4 (en) 2021-07-21
SG11201910115XA (en) 2019-11-28

Similar Documents

Publication Publication Date Title
US20240041914A1 (en) RNAi APPROACH FOR CROP PEST PROTECTION
Song et al. MicroRNAs and their regulatory roles in plant–environment interactions
JP6223187B2 (en) Nucleic acid molecules that confer resistance to Coleoptera
RU2653752C2 (en) Down-regulating gene expression in insect pests
Xue et al. New approaches to agricultural insect pest control based on RNA interference
RU2665549C1 (en) Down-regulating gene expression in insect pests
JP2014503218A (en) A nucleic acid molecule that targets Rho1 small GTP-binding protein and confers resistance to Coleoptera pests
CN105473605A (en) Phi-4 polypeptides and methods for their use
JP2014504863A (en) Nucleic acid molecules that target the C subunit of vacuolar ATPase and confer resistance to Coleoptera pests
TW201623613A (en) DRE4 nucleic acid molecules that confer resistance to coleopteran pests
TW201321508A (en) Nucleic acid molecules that target RPA70 and confer resistance to coleopteran pests
CN112770761A (en) Use of RNA molecules for controlling plant pests
JP2017510246A (en) RnapII-140 nucleic acid molecule conferring resistance to Coleoptera
JP2018513675A (en) RNA polymerase II33 nucleic acid molecules for controlling pests
Ren et al. Assessment on effects of transplastomic potato plants expressing Colorado potato beetle β-actin double-stranded RNAs for three non-target pests
US20170349907A1 (en) Methods to Monitor Post-Translational Gene Silencing Activity in Plant Tissues/Cell Types Relevant for Pathogen Entry, Propagation or Replication
JP2018524004A (en) PRP8 nucleic acid molecules for controlling pests
Maligeppagol et al. Transgenic tomato expressing dsRNA of juvenile hormone acid O-methyl transferase gene of Helicoverpa armigera (Lepidoptera: Noctuidae) affects larval growth and its development
CN112204141A (en) Short/small hairpin RNA molecules
CN110637092A (en) Use of structure specific recognition protein 1(SSRP1) nucleic acid molecules for controlling insect pests
OA20111A (en) Rnai approach for crop pest protection.
Chatterjee et al. Amenability of Maruca vitrata (Lepidoptera: Crambidae) to gene silencing through exogenous administration and host-delivered dsRNA in pigeonpea (Cajanus cajan L.)
US20220162626A1 (en) Modulation of Transgene Expression
Jaiwal et al. Host-induced RNA interference confers insect resistance in Tobacco by targeting chitin synthase gene of Helicoverpa armigera
Khabbazi et al. Different Generations of Genetically Modified Crops for Insect Resistance